人脂肪干细胞的获取及其定向分化为内皮细胞差异蛋白筛选的实验研究
|
摘要
为解决血管组织工程种子细胞来源不足问题,及探索干细胞定向内皮分化的分子机制,本实验以人脂肪干细胞(hADSCs)为研究对象,进行了hADSCs体外分离、培养、扩增和生物学特性的鉴定,最佳内皮细胞诱导分化体系的探索及应用蛋白质组学技术从蛋白质角度筛选hADSCs内皮分化过程中所表达的差异蛋白。
应用胶原酶消化和贴壁筛选联合培养的方法,从人脂肪组织中快速分离获得了hADSCs,经一般生物学特性的检测,证实了我们获得的hADSCs具有较高的纯度,能够稳定的进行传代培养以及冻存、复苏,这符合组织工程对种子细胞的要求。从细胞因子与细胞外基质的角度探讨其对hADSCs定向内皮分化的影响,最终摸索出以纤维粘连蛋白(FN)作为细胞外基质与富含多种生长因子的内皮细胞支持液EGM2-MV﹢高浓度VEGF165(50ng/ml)组成的诱导条件,构成了hADSCs最佳内皮细胞诱导分化的体系。应用蛋白质组学技术首次从蛋白质角度筛选出了hADSCs定向分化为内皮细胞的差异蛋白。为进一步了解其分化过程中的调控机制提供了重要的实验基础。
综上所述,本研究建立了hADSCs体外分离培养及大规模扩增的简易技术;建立了hADSCs体外最佳的内皮细胞诱导分化体系;筛选出hADSCs内皮分化过程中的差异蛋白,初步分析发现了内皮分化相关的10个重要蛋白。上述研究结果为探索hADSCs定向分化的分子机制提供了理论基础,也为人们更好地将hADSCs应用于临床治疗缺血性疾病或血管依赖性疾病,促进组织工程化器官的血管再生,提供了充分的实验依据。
With the increasing incidence of vascular diseases, the desire for the vascular transplantation materials is also continually expanded. In order to solve a variety of problems about the synthetic materials and the autologous vascular transplantation, scientists tried many ways to construct tissue-engineered blood vessels. And how to obtain the‘seeding cells’is one of the challenges for us.Up to now,some achievements has been got in the treatment of ischemic diseases with endothelial cells. Considering the difficulty in obtaining and amplifying endothelial cells,seeking for suitable seeding cells to construct tissue-engineered blood vessels has been a hot pot. The stem cells have been the favorite choice for seeding cells in tissue engineering due to highly proliferative ability and multipotentiality.
Nowadays, there are two kinds of stem cells: embryonic stem cells and adult stem cells. Due to the limitation of embryonic stem cells,such as ethical problems and inconvenient harvest, adult stem cells have gradually become investigative focus. Compared with bone marrow-derived mesenchymal stem cells, ADSCs(Human adipose-derived stem cells) are also multipotent stem cells derived from the mesoderm, which possess potent proliferative capacity and self-renewal and multilineage differentiation.But ADSCs have some merit, including drawing the materials easily, slightly injury, multiple amplification in vitro and induced differentiation easily, which make them become the main source of tissue engineered vascular seeding cells.
In 2001, Zuk[27]obtained adipose-derived stem cells from human adipose tissue. Subsequently scientists modified several methods. So far,there is no acceptable, simple or efficient method for isolating and cultivating highly homogenous and undifferentiated hADSCs. Moreover, methods for identification have not been fully established. Therefore, it is inevitable to establish highly efficient method to isolate and identify hADSCs.
The molecular mechanism of hADSCs directional differentation into endothelial cells is not clear. Only the mechanism of differentiated regulation are found out, it will offer abundant seeding cells for tissue engineering. Because both the stem cell research and proteomics research are hot spots in life sciences in the 21th century, if we want to recognize the complex activities of stem cells, we must focus on the proteins from the whole and dynamic point of view.
In the present study, hADSCs were isolated, cultivated and amplified,and the biological characteristics of hADSCs were identified as well. Then hADSCs were induced into endothelial cells by utilizing cytokine and extracellular matrix. Furthermore, hADSCs differential proteins in directional differentiation into endothelial cells were screened by proteomics techniques.
The results show that we have successfully isolated and cultured highly homogenous and undifferentiated hADSCs, which have multilineage differentiation properties. The most important finding is that one gram of adipose tissue can yield～1×10~6 adipose-derived mesenchymal stem cells, equivalent to the yield from 40ml bone marrow. In addition,hADSCs are negative for histocompatibility complex molecules (HLA-DR), which implies that hADSCs are suitable for allograft without MHC limitation. hADSCs we have got can be passaged, freezed and revived stably. These characteristics will better meet the demands of cell transplant and tissue engineering.
The multi-differentiation ability is the most important identification for stem cells[20]. The results show that human adipose-derived mesenchymal stem cells has potentiality to be induced into adipocytes, osteocytes and endothelial cells. In this experiment, we compared the differentiation process at different time points. With the extension of time, the capacity of being induced into adipocytes,or osteocytes is also increased.
We study the factors which impact hADSCs directional differentiation into endothelial from the cytokine and extracellular matrix point of view,and find that the combination of FN as extracellular matrix and endothelial cell supporting culture medium EGM2-MV as well as high concentration VEGF165(50ng/ml)is optimal for endothelial differentiation.
We screened directional differentially expressed proteins of in hADSCs differentiation into endothelial cells by using proteomic techniques, then we identifed these proteins by using MALDITOF -TOF, and we got 10 key regulatory proteins related to the differentiation of hADSCs into endothelial cells, which provides an important experimental foundation for understanding the differentiation process in the regulatory mechanism.
In conclusion, we have successfully isolated and cultured highly homogenous and undifferentiated hADSCs, optimized the induced differentiation system of endothelial cells, and for the first time in this experiment, we screened directional differentially expressed proteins in hADSCs differentiation into endothelial cells by proteomic technologies. The results provides not only a theoretical basis for the molecular mechanism of hADSCs induced differentiation into endothelial cells, but also a sufficient experimental evidence in using hADSCs to treat ischemic diseases or vascular-dependent diseases and promote blood vessel regeneration in tissue-engineered organs.
应用胶原酶消化和贴壁筛选联合培养的方法,从人脂肪组织中快速分离获得了hADSCs,经一般生物学特性的检测,证实了我们获得的hADSCs具有较高的纯度,能够稳定的进行传代培养以及冻存、复苏,这符合组织工程对种子细胞的要求。从细胞因子与细胞外基质的角度探讨其对hADSCs定向内皮分化的影响,最终摸索出以纤维粘连蛋白(FN)作为细胞外基质与富含多种生长因子的内皮细胞支持液EGM2-MV﹢高浓度VEGF165(50ng/ml)组成的诱导条件,构成了hADSCs最佳内皮细胞诱导分化的体系。应用蛋白质组学技术首次从蛋白质角度筛选出了hADSCs定向分化为内皮细胞的差异蛋白。为进一步了解其分化过程中的调控机制提供了重要的实验基础。
综上所述,本研究建立了hADSCs体外分离培养及大规模扩增的简易技术;建立了hADSCs体外最佳的内皮细胞诱导分化体系;筛选出hADSCs内皮分化过程中的差异蛋白,初步分析发现了内皮分化相关的10个重要蛋白。上述研究结果为探索hADSCs定向分化的分子机制提供了理论基础,也为人们更好地将hADSCs应用于临床治疗缺血性疾病或血管依赖性疾病,促进组织工程化器官的血管再生,提供了充分的实验依据。
With the increasing incidence of vascular diseases, the desire for the vascular transplantation materials is also continually expanded. In order to solve a variety of problems about the synthetic materials and the autologous vascular transplantation, scientists tried many ways to construct tissue-engineered blood vessels. And how to obtain the‘seeding cells’is one of the challenges for us.Up to now,some achievements has been got in the treatment of ischemic diseases with endothelial cells. Considering the difficulty in obtaining and amplifying endothelial cells,seeking for suitable seeding cells to construct tissue-engineered blood vessels has been a hot pot. The stem cells have been the favorite choice for seeding cells in tissue engineering due to highly proliferative ability and multipotentiality.
Nowadays, there are two kinds of stem cells: embryonic stem cells and adult stem cells. Due to the limitation of embryonic stem cells,such as ethical problems and inconvenient harvest, adult stem cells have gradually become investigative focus. Compared with bone marrow-derived mesenchymal stem cells, ADSCs(Human adipose-derived stem cells) are also multipotent stem cells derived from the mesoderm, which possess potent proliferative capacity and self-renewal and multilineage differentiation.But ADSCs have some merit, including drawing the materials easily, slightly injury, multiple amplification in vitro and induced differentiation easily, which make them become the main source of tissue engineered vascular seeding cells.
In 2001, Zuk[27]obtained adipose-derived stem cells from human adipose tissue. Subsequently scientists modified several methods. So far,there is no acceptable, simple or efficient method for isolating and cultivating highly homogenous and undifferentiated hADSCs. Moreover, methods for identification have not been fully established. Therefore, it is inevitable to establish highly efficient method to isolate and identify hADSCs.
The molecular mechanism of hADSCs directional differentation into endothelial cells is not clear. Only the mechanism of differentiated regulation are found out, it will offer abundant seeding cells for tissue engineering. Because both the stem cell research and proteomics research are hot spots in life sciences in the 21th century, if we want to recognize the complex activities of stem cells, we must focus on the proteins from the whole and dynamic point of view.
In the present study, hADSCs were isolated, cultivated and amplified,and the biological characteristics of hADSCs were identified as well. Then hADSCs were induced into endothelial cells by utilizing cytokine and extracellular matrix. Furthermore, hADSCs differential proteins in directional differentiation into endothelial cells were screened by proteomics techniques.
The results show that we have successfully isolated and cultured highly homogenous and undifferentiated hADSCs, which have multilineage differentiation properties. The most important finding is that one gram of adipose tissue can yield～1×10~6 adipose-derived mesenchymal stem cells, equivalent to the yield from 40ml bone marrow. In addition,hADSCs are negative for histocompatibility complex molecules (HLA-DR), which implies that hADSCs are suitable for allograft without MHC limitation. hADSCs we have got can be passaged, freezed and revived stably. These characteristics will better meet the demands of cell transplant and tissue engineering.
The multi-differentiation ability is the most important identification for stem cells[20]. The results show that human adipose-derived mesenchymal stem cells has potentiality to be induced into adipocytes, osteocytes and endothelial cells. In this experiment, we compared the differentiation process at different time points. With the extension of time, the capacity of being induced into adipocytes,or osteocytes is also increased.
We study the factors which impact hADSCs directional differentiation into endothelial from the cytokine and extracellular matrix point of view,and find that the combination of FN as extracellular matrix and endothelial cell supporting culture medium EGM2-MV as well as high concentration VEGF165(50ng/ml)is optimal for endothelial differentiation.
We screened directional differentially expressed proteins of in hADSCs differentiation into endothelial cells by using proteomic techniques, then we identifed these proteins by using MALDITOF -TOF, and we got 10 key regulatory proteins related to the differentiation of hADSCs into endothelial cells, which provides an important experimental foundation for understanding the differentiation process in the regulatory mechanism.
In conclusion, we have successfully isolated and cultured highly homogenous and undifferentiated hADSCs, optimized the induced differentiation system of endothelial cells, and for the first time in this experiment, we screened directional differentially expressed proteins in hADSCs differentiation into endothelial cells by proteomic technologies. The results provides not only a theoretical basis for the molecular mechanism of hADSCs induced differentiation into endothelial cells, but also a sufficient experimental evidence in using hADSCs to treat ischemic diseases or vascular-dependent diseases and promote blood vessel regeneration in tissue-engineered organs.
引文
[1]Langer R,Vacanti JP. Tissue engineering[J]. Science,1993,260(5110):920-926.
[2]NIH. Stem Cells : Scientific Progress and Future Research Directions.(August 6 ,2001) Website : http :/ / www. nih. g ov/ news/ stemcell/ scireport . htm
[3]Hochedlinger K,Jaenisch R.Nuclear reprogramming and pluripotency [J]. Nature, 2006,441(7097):1061-1067.
[4]Rodolfa KT, Eggan K.A transcriptional logic for nuclear reprogramming [J].Cell,2006,126(4):652-655.
[5]Cervera RP,Stojkovic M.Commentary:somatic cell nuclear transfer-progress and promise[J].Stem Cells,2008,26(2):94- 495.
[6]Daley GQ, Goodell MA, Snyder EY. Realistic prospects for stem cell therapeutics[R].Hematology Am Soc Hematol Educ Program,2003: 398-418.
[7]Evans M J, Kaufman M H. Establishment in culture of pluripotent cells from mouse embryos[J]. Nature,1981,292(9):154-156.
[8]Takahashi K,Yamanaka S.Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors[J]. Cell, 2006, 126(4):663-676.
[9]Hyun I,Hochedlinger K, Jaenisch R,et al. New advances in iPS cell research do not obviate the need for human embryonic stem cells[J].Cell Stem Cell, 2007,1(4): 367-368.
[10]Watts G. Scientists“reprogramme”skin cells to create embryonic stem cells[J]. BMJ, 2007,335(7630): 1112-1113.
[11]Cyzy J, Wiese C, Rolletschek A,et al.Potential of embryonic and adult stem cells in vitro[J]. Biol Chem,2003,384(10-11):1391-1409.
[12]程冬婉,张伟.成体干细胞临床应用的研究[J].中国肿瘤生物治疗杂志,2006,13(2):150-152.
[13]周长梅.干细胞研究进展[J].淮北煤炭师范学院学报,2006,27(4):45-48.
[14]Watts C, McConkey H, Anderson L,et al.Anatomical perspectives onadultneural stem cells[J]. JAnat, 2005, 207(3): 197-208.
[15]SchafflerA, BuchlerC.Concise review: adipose tissue-derived stromal cells--basic and clinical implications fornovel cell-based therapies[J]. Stem Cells, 2007,25(4): 818-827.
[16]Mimeault M, Hauke R,Mehta PP,etal.Recent advances in cancer stem /progenitor cell research: therapeutic implications for overcoming resistance to the most aggressive cancers[ J]. J Mol Cell Med, 2007, 11(5): 981-1011.
[17]Minguell JJ, Er ices A,Conget P. Mesenchymal stem cells[J]. Exp Biol Med, 2001, 226 (6) : 507-520.
[18]曹谊林,崔磊,刘伟.组织工程学理论与实践[M].北京:科学出版社,2008年,第一版.
[19]Ng AM, Kojima K, Kodoma S, et al. Isolation techniques of murine bone marrow progenitor cells and their adipogenic, neurogenic and osteogenic differentiation capacity[J]. Med J Malaysia. 2008,63:121-122.
[20]Zuk PA, Zhu M, Ashjian P, et al.Human adipose tissue is a source of multipotent stem cells[J]. Mol Biol Cell 2002,13: 4279–4295
[21]Gronthos S, Franklin DM, Leddy HA, et al. Surface proteincharacterization of human adipose tissue- derived stromal cells [J]. J Cell Physiol, 2001, 189: 54- 63.
[22]Safford KM, Hicok KC, Safford SD, et al. Neurogenic differentiation of urine and human adipose- derived stromal cells [J]. Biochem Biophys Res Commun,2002,294:371- 379.
[23]Miranville A, Heeschen C, Sengenes C, et al. Improvement of postnatal neovascularization by human adipose tissue- derived stem cells [J]. Circulation, 2004,20:349- 355.
[24]Planat- Benard V, Silvestre JS, Cousin B, et al. Plasticity of human adipose lineage cells toward endothelial cells:physiological and therapeutic perspec- tives[J]. Circulation,2004,10:656- 663.
[25]De Ugarte DA, Morizono K, Elbarbary A, et al.Comparison of multi-lineage cells from human adipose tissue and bone marrow[J]. Cells Tissues Organs 2003,174: 101-109.
[26]Izadpanah R, Trygg C, Patel B, et al. Biologic properties of mesenchymal stem cells derived from bone marrow and adipose tissue[J].J Cell Biochem, 2006,99:1285-1297.
[27]Zuk PA, Zhu M, Mizuno H, et al. Multilineage cells from human adipose tissue: implications for cell-based therapies[J].Tissue Eng 2001,7: 211–228.
[28]Mizuno H, Hyakusoku H. Mes engenic potential and future clinical perspective of human proces sed lipoaspirate cel ls [J]. J Nippon Med Sch, 2003,70(4):300- 306.
[29]金杰,虞渝生.脂肪干细胞研究进展.浙江医学.2008,30(1):101-103.
[30]Lee RH,Kim B,Choi I,etal.Characterization and expression analysis ofmesenchymal stem cells from human bonemarrow and adipose tissue [J]. Cell Physiol Biochem, 2004,14(4/6): 311-324.
[31]Gimble J, Guilak F. Adipose-derived adult stem cells: isolation, characteri- zation,and differentiation potential [J]. Cytotherapy, 2003,5(5): 362-369.
[32] Guilak F, Lott KE, Awad HA, et al. Clonal Analysis of the Differentiation Potential of Human Adipose-Derived Adult Stem Cells[J].J Cell Physiol, 2006,206: 229–237.
[33]Mi Hyang Moon, Sun Young Kim, Yeon Jeong Kim1,et al. Human Adipose Tissue-Derived Mesenchymal Stem Cells Improve Postnatal Neovascula- rization in a Mouse Model of Hind limb Ischemia[J]. Cell Physiol Biochem, 2006,17:279-290.
[34]Ying Cao, Zhao Sun, Lianming Liao,et al. Human adipose tissue-derived stem cells differentiate into endothelial cells in vitro and improve postnatal neovascularization in vivo[J]. Biochemical and Biophysical Research Communications,2005,332:370–379.
[35]E. Raposio, C. Guida, I. Baldelli,et al. Characterization and induction ofhuman pre-adipocytes[J]. Toxicology in Vitro,2007, 21:330–334.
[36]Naoki Yamamoto, Hirohiko Akamatsu, Seiji Hasegawa,et al. Isolation of multipotent stem cells from mouse adipose tissue[J]. Journal of Dermat- ological Science,2007,48:43-52.
[37]De Ugarte DA, Alfonso Z, Zuk PA,et al. Differential expression of stem cell mobilization-associated molecules on multi-lineage cells from adipose tissue and bone marrow[J].Immunol Lett,2003,89:267–270.
[38]Lin TM, Tsai JL, Lin SD,et al. Accelerated growth and prolonged lifespan of adipose tissue-derived human mesenchymal stem cells in a medium using reduced calcium and antioxidants[J].Stem Cells Dev, 2005,14(1):92–102.
[39]Tholpady SS, Llull R, Ogle RC,et al.Adipose tissue: stem cells and beyond [J].Clin Plast Surg,2006,33(1):55–62.
[40]Gimble JM. Adipose tissue-derived therapeutics[J].Expert Opin Biol Ther, 2003,3(5):705–13.
[41]Cheryl T. Gomillion, Karen J.L. Burg. Stem cells and adipose tissue engineering[J].Biomaterials,2006,27: 6052–6063.
[42]Katz A J, Ashok T, Sunil S T,et al. Cell surface and transcriptional characterization of human adipose-derived adherent stromal(hADAS) cells [J]. Stem Cells,2005,23:412-423.
[43]Strem BM, Hicok KC, Zhu M,et al. Multipotential differentiation of adipose tissue-derived stem cells[J]. Keio J Med,2005,54(3):132-141.
[44]郭新,赵东海,何旭等.大鼠骨髓间充质干细胞的分离培养[J].吉林大学学报:医学版,2005,31(3):469-471.
[45]崔磊,尹烁,杨平等.脂肪干细胞HLA分子表达与体外抑制淋巴细胞增殖的实验研究[J].中华医学杂志,2005,85(27):1890-1894.
[46]Aust L, Devlin B, Foster SJ,et al.Yield of human adiposederived adult stem cells from liposuction aspirates[J].Cytotherapy, 2004,6:7-14.
[47]Lee RH, Kim B, Choi I,et al.Characterization and expression analysis ofmesenchymal stem cells from human bone marrow and adipose tissue[J].Cell Physiol Biochem,2004,14:311-324.
[48]Planat-Benard V, Silvestre JS, Cousin B,et al. Plasticity of human adipose lineage cells toward endothelial cells: physiological and therapeutic perspectives[J].Circulation,2004,109:656–663.
[49]Rangappa S, Fen C, Lee EH,et al.Transformation of adult mesenchymal stem cells isolated from the fatty tissue into cardiomyocytes[J]. Ann Thorac Surg,2003,75:775-779.
[50]Safford KM, Hicok KC, Safford SD,et al.Neurogenic differentiation of murine and human adipose-derived stromal cells[J].Biochem Biophys Res Commun,2002,294:371-379.
[51]Ying Cao, Zhao Sun, Lianming Liao,et al. Human adipose tissue -derived stem cells differentiate into endothelial cells in vitro and improve postnatal neovascularization in vivo[J]. Biochemical and Biophysical Research Communications,2005,332:370-379.
[52]Strem BM, Hicok KC, Zhu M,et al. Multipotential differentiation of adipose tissue-derived stem cells[J]. Keio J Med, 2005,54:132-141.
[53]Slaek,JM. Stem ells in Epithelial Tissues[J].Science,2000,287: 1431-1433.
[54]He X, Li YL, Wang XR,et al. Mesenchymal stem cells transduced by PLEGFP-N1 retroviral Vector maintain their biological features and differentiation[J].Chin Med J,2005,118:1728-1734.
[55]李慧武,戴赶戎,汤亭亭等.长期体外培养对人脂肪源性间充质干细胞生物学特性的影响[J].中华创伤骨科杂志,2006,8(10):929-932.
[56]Bartholomew A,Sturgeon C,Siatskas M,et al.Mesenehymal stem cells suppress lymphocyte Proliferation in vitro and Prolong skin graft survival in vivo[J].Exp Hematol,2002,30:42-48.
[57]Ogawa R, Mizuno H, Hyakusoku H,et al. Chondrogenic and osteogenic differentiation of adipose derived stem cells isolated from GFP transgenicmice[J]. J Nippon Med Sch,2004,71(4):240-241.
[58]Huang J I, Zuk P A, Jones N F,et al. Chondrogenic potential of multipotential cells from human adipose tissue[J]. Plast Recons trSurg,2004, 113(2): 585- 594.
[59]杨亚军,朱庆生. CDMPI诱导大鼠脂肪干细胞体外成软骨细胞的实验[J].第四军医大学学报,2008,29(4):342-345.
[60]陈崎,彭智.脂肪干细胞在骨、软骨组织工程中的研究进展[J].中国民族民间医药.2009,14:26-27.
[61]Liu Y, LuoE, Chen X,et al. Molecular and cellular characterization during chondrogenic differentiation of adipose tissue-derived stromal cells in vitro and cartilage formarionin vivo[J]. J Cell Mol Med,2005,9(4):929-939.
[62]Mochizuki T, Muneta T, SakaguchiY,et al. Higher chondrogenic potentialof fibrous synovium-and adipose synovium-derived cells compared with subcutaneous fat-derived cells:Distinguishingproperties ofmesenchymal stem cells in humans[J]. ArthritisRheum,2006,54(3):843-853.
[63]SegawaY, MunetaT, MakinoH,et al. Mesenchymal stem cells derived from synovium, meniscus, anterior cruciate ligament,and articular chondrocytes share similar gene expression profiles[J]. J Orthop Res,2009,27(4):435-441.
[64]Huang J I, Zuk P A, Jones N F, et al. Chondrogenic potential of multipotential cells from human adipose tissue[J]. Plast Recons trSurg, 2004, 113(2):585-594.
[65]杨平,尹烁,崔磊等.脂肪干细胞向血管平滑肌细胞诱导的实验研究[J].中国修复重建外科杂志,2008,22:481-486.
[66]张端珍,盖鲁粤,刘宏伟等.脂肪干细胞诱导成心肌细胞研究[J].心脏杂志,2005,17:405-408.
[67]Mizuno H, Zuk PA, Zhu M,et al. Myogenic differentiation by human processed lipoaspirate cells[J]. Plast Reconstr Surg,2002,109(1): 199-209.
[68]何旭,王心蕊,张海英等.人骨髓间充质干细胞体外定向诱导分化为脂肪细胞方法的建立[J].吉林大学学报,2006,3(32):530-532.
[69]Herrera-Herrera M, Zapata-Bustos R, Salazar-Olivo L. Simplified culture techniques for growth and differentiation of murine and human pre-adipocytes for translational applications[J]. Cytotherapy, 2009, 11(1): 52-60.
[70]张勇.脂肪干细胞的研究进展[J].医学研究生学报,2009,22(10):1088- 1092.
[71]Park JP, Jung JW, Lee YS,et al. The roles of Wnt antagonists Dkk1 and sFRP4 during adipogenesis of human adipose tissue-derived mesenchymal stem cells[J]. Cell Prolif,2008, 41(6):859-874.
[72]Scheideler M, Elabd C, Zaragosi LE,et al.Comparative transcriptomics of human multipotent stem cells during adipogenesis and osteoblastogenesis [J].BMC Genomics,2008, 17(9):340.
[73]Chen J, Zhang ZG, Ly Y,et al.Intravenousadministration of human bone marrow stromal cells induces angiogenesis in the ischemic boundary zone after stroke in rats[J].Circ Res,2003,92(6):692-699.
[74]Haynes worth SE, Baber MA, Caplan AI. Cell surface antigens on human marr ow-derived mesenchymal cells are detected by monoclonal antibodies [J]. Bone,1992,13:68-69.
[75]Barry FP,Boynton RE,Haynesworth S,et al.The monoclonal antibody SH-2,raised against human mesenchymal stem cells,recognizes an epitope on endoglin(CD105)[J].Biochem Biophys Res Commun, 1999,265(1):134-139.
[76]Fleming JE,Haynesworth SE,Cassiede P,et al.Monoclonal antibody against adult marrow-derived mesenchymal stem cells recognizes developing vasculature in embryonic human skin[J].Dev Dyn,1998,212(1):119-132.
[77]Ohneda O,Ohneda K,Arai F,et al.ALCAM(CD166):its role in hematopoietic and endothelial development[J].Blood,2001,98 (7):2134-2142.
[78]Reyes M,Verfaillie CM.Characterization of multipotent adult progenitor cells,a subpopulation of mesenchymal stem cells[J].Ann NY Acad Sci, 2001,938:231-235.
[79]Reyes M,Dudek A,Jahagirdar B,et al.Origin of endothelial progenitors in human postnatal bone marrow[J].J Clin Invest,2002,109(3):337-346.
[80]Lozito TP, Kuo CK, Taboas JM,et al. Human Mesenchymal Stem Cells Express Vascular Cell Phenotypes Upon Interaction With Endothelial Cell Matrix[J]. Cellular Biochemistry,2009,107:714–722.
[81] Oswald J, Boxberger S, J?rgensen B,et al. Mesenchymal Stem Cells Can Be Differentiated Into Endothelial Cells In Vitro[J].STEM CELLS.2004,22: 377- 384.
[82]Chen MY, Lie PC, Li ZL,et al. Endothelial differentiation of Wharton's jelly–derived mesenchymal stem cells in comparison with bone marrow–derived mesenchymal stem cells[J].Exp Hematol,2009,37: 629-640.
[83]Valérie Planat-Benard, Jean-Sébastien Silvestre, Béatrice Cousin,et al. Plasticity of Human Adipose Lineage Cells Toward Endothelial Cells Physiological and Therapeutic Perspectives[J].Circulation,2004,109:656- 663.
[84] De Francesco F, Tirino V, Desiderio V,et al. Human CD34+/CD90+ASCs Are Capable of Growing as Sphere Clusters, Producing High Levels of VEGF and Forming Capillaries[J].PLoS ONE,2009,4(8):1-13.
[85]Ying Cao, Zhao Sun, Lianming Liao,et al.Human adipose tissue-derived stem cells differentiate into endothelial cells in vitro and improve postnatal neovascularization in vivo[J]. Biochemical and Biophysical Research Communications,2005,332: 370-379.
[86]Watt F,Hogan B.Out of Eden:stem cells and their niches[J]. Science,2000, 287(5457):1427-1430.
[87]Roberts DM,Kearney JB,Johnson JH,et al.The vascular endothelial growth factor(VEGF) receptor Flt-1(VEGFR-1) modulates Flk-1(VEGFR-2) signaling during blood vessel formation[J].Am J Pathol,2004, 164(5): 1531- 1535.
[88]Kearney JB,Kappas NC,Ellerstrom C,et al.The VEGF receptor flt-1 (VEGFR-1) is a positive modulator of vascular sprout formation and branching morphogenesis[J].Blood,2004,103(12):4527-4535.
[89]Wang M,Crisostomo PR,Herring C,et al.Human progenitor cells from bone marrow or adipose tissue produce VEGF,HGF,and IGF-I in response to TNF by a p38 MAPK-dependent mechanism[J].Am J Physiol Regul Integr Comp Physiol,2006,291(4):880-884.
[90]Zeng Q,Li X,Beck G,et al.Growth and differentiation factor-5(GDF-5) stimulates osteogenic differentiation and increases vascular endothelial growth factor(VEGF) levels in fat derived stromal cells in vitro[J].Bone, 2007,40(2):374-381.
[91]Crisostomo PR,Wang M,Herring CM,et al.Gender differences in injury induced mesenchymal stem cell apoptosis and VEGF,TNF,IL-6 expression: Role of the 55 kDa TNF receptor(TNFR1)[J].J Mol Cell Cardiol, 2007,42(1):142-149.
[92]Oswald J,Boxberger S,Jorgensen B,et al.Mesenchymal stem cells can be differentiated into endothelial cells in vitro[J].Stem Cells,2004,22(3): 377- 384.
[93]Al-Khaldi A,Eliopoulos N,Martineau D,et al.Postnatal bone marrow stromal cells elicit a potent VEGF-dependent neoangiogenic response in vivo[J].Gene Ther,2003,10(8):621-629.
[94]Annabi B,Naud E,Lee YT,et al.Vascular progenitors derived from murine bone marrow stromal cells are regulated by fibroblast growth factor and are avidly recruited by vascularizing tumors[J].J Cell Biochem,2004,91(6): 1146-1158.
[95]Adams JC,Watt FM. Regulation of development and differentiation by the extra cellular matrix[J].Development,1993,117(4):1183- 1198.
[96]郭新,李伟,朱桂彬等.基质蛋白与整合素参与人骨髓间充质干细胞体外内皮的分化[J].中国临床康复,2005,9(14):58-60.
[97]Meredith JE,Fazeli B,Schwartz MA.The extracellular matrix as a cell survival factor[J].Mol Biol Cell,1993,4(9):953-961.
[98]Annabi B,Lee YT,Turcotte S,et al. Hypoxia promotes murine bone- marrow- derived stromal cell migration and tube formation[J].Stem Cells,2003,21 (3):337-347.
[99]Remy-Zolghadri M. Endothelium ProPerties of a tissue-engineered blood Vessel for small-diameter vascular reconstruetion[J].J Vase Surs,2004,39(3): 613-620.
[100]Dejana,E.Endothelial cell-cell junctions:happy together[J].Nat Rev Mol Cell Biol,2004,5(4):261-270.
[101]Feraud,0,Y.Cao,D.Vittet.Embryonic stem cell-derived embryoid bodies development in collagen gels recapitulates sprouting angiogenesis[J].Lab Invest,2001,81(12):1669-1681.
[102]Feraud,0.,M.H.Prandini,D.Vittet,Vaseulogenesis and angiogenesis from in vitro differentiation of mouse embryonic stem cells[J]. Methods Enzymol, 2003,365:214-228.
[103]Mallet,C.TGFbetal induces vaseulogenesis and inhibits angiogenic Sprout- ing in an embryonic stem cell differentiation model: respeetive Contribution of ALK1 and ALK5[J].Stem Cells.2006, 24(11):2420-2427.
[104]Lyden,D.Impaired recruitment of bone-marrow-derived endothelial and he- matopoietic Precursor cells blocks tumor angiogenesis and growth[J].Nat Med,2001,7(11):1194-1201.
[105]Wikins MR,Pasquali C.From Proteins to Proteomes:Large scalc protein identification by two-dimensional electrophoresis and amino acid analysis [J].Bio/Technology,1996,14:6561-6567.
[106]Petricoin EF,Zoon KC.Clinical proteomics:translating benchside promise into bedside reality[J].Nature Rev Drug Discov,2002, 1(9):683-695.
[107]Petricoin EF,Zoon KC.Clinical proteomics:translating benchside promise into bedside reality[J].Nature Rev Drug Discov,2002,1(9):683-695.
[108]P.Kahn,From genome to Proteome:looking at a cell proteins[J]. Science, 1995,270:369-370.
[109]D.Swinbanks,Govenmment backs proteome proposal[J].Nature, 1995,378: 653.
[110]Beranova G,Iorgianni S.Proteome analysis by two-dismensional gel electrophoresis and mass spectrometry:strengths and limitations [J].Trends in Analytical Chemistry,2003,22(5): 273-280.
[111]O Parrell PH.High resolution two-dimensional elcetrophoresis of proteins [J].Journal of Biological Chemistry,1975,250:4007 -4021.
[112]Li C,Tan YX,Zhou H,et al. Proteomic analysis of hepatitis B virus- associated hepatocellular carcinoma:Identification of Potential tumor markers[J].Proteomics,2005,5(4):1125-1139.
[113]Sun W,Xing B,Sun Y,et al. Proteome analysis of hepatocellular carcinoma by two-dimensional Difference gel electrophoresis: novel protein markers in hepatocellular carcinoma tissues[J]. MOL Cell Proteomics,2007,6(10): 1798-1808.
[114]Blanc JF,Lalanne C,Plomion C,et al.Proteomic analysis of Differentially expressed Proteins in hepatocellular carcinoma developed in Patients with chronic viral hepatitis C[J]. Proteomics,2005,5(14):3778-3789.
[115]Kang SM,Shin MJ,Kim JH,et al.Proteomic Profiling of cellular Proteins Interacting with the hepatitis C virus core Protein[J]. Proteomics,2005,5(8): 2227-2237.
[116]Liang CR,Leow CK,Neo JC,et al.Proteome analysis of human hepatocell ular carcinoma tissues by two-dimensional difference gel electrophoresis and mass spectrometry[J]. Proteomics,2005, 5(8):2258-2271.
[117]Jorgenson JW,Lukacs KD.Zone electrophoresis in open-tubular glasscapillareies[J].Anal Chem,1981,53:1298.
[118]Garcia Campana AM,Gamiz Gracia L,Baeyens WR,et al.Dervatization of bicmolecules for chemilum inescent detection in capillary electrophor esis[J]. Chromatogr B Analyt Technol Bicmed Life Sci,2003,793:49-74.
[119]Shevchenko A,Loboda A,Shevchenko A,et al. MALDI quadrupole time-of– flight mass spectrometry:a powerful tool for proteomic research[J].Anal Chem,2000,72:2132-2141.
[120]Minden J,Waggoner A.[R].US patent:6043025,2000.
[121]Venter JC,Adams MD,Myers EW,et al.The sequence of the humangen- ome[J].Science,2001,291(5507):1304-1351.
[122]Mike Tyers,Matthias Mann M.From genomics to Proteomics[J]. Nature, 2003,422(6928):193-197.
[123]Pandey A,Manll M.Proteomics to study genes and genomes[J].Nature, 2000,405(6788):837-846.
[124]Ruedi Aebersold,Matthias Mann. Mass spectrometry-based Proteomics [J]. Nature,2003,422(6928):198-207.
[125]Ong SE,Mann M.Mass spectrometry-based Proteomics turns quantitative [J].Nature,Chemical Biology,2005,l(5):252-262.
[126]Zvonic S,Lefevre M, Kilroy G,et al.Secretome of primary cultures of human adipose-derived stem cells:modulation of serpins by adipogenesis[J].Mol Cell Proteomics,2007,6 (1):18-28.
[127]Pittenger MF, Mackay AM, Beck SC,et al. Multilineage potential of adult human mesenchymal stem cells[J]. Science.1999, 284: 143–147.
[128]李文婷,孙华林,许增禄等。大鼠骨髓基质细胞向Schwann细胞分化后蛋白表达变化的研究[J].中国科学,2009,39(2):167-178.
[129]Wang D,Gao L.Proteomic analysis of neural differentiation of mouse embryonic stem cells[J].Proteomics,2005,5(17):4414- 4426.
[130]Elliott ST,Crider DG, Garnham CP,et al. Two-dimensional gel electrop-horesis database of murine R1 embryonic stem cells[J].Proteomics, 2004,4(12):3813-3832.
[131]Wang J,RaoS,Chu J,etal.A protein interaction network for pluripotency of embryonic stem cells[J].Nature,2006,444(7117): 364-368.
[132]Prudhomme W,Daley GD, Zandstra P,etal.Multivariate proteomic analysis ofmurine embryonic stem cel self-renewal versus differentiaion signaling[J]. ProcNatlAcad SciUSA, 2004, 101(9): 2900-2905.
[133]Baharvand H, Fathi A, Gourabi H,et al. Identification of mouse embryonic stem cell-associated proteins[J]. JProteomeRes, 2008,7(1): 412-423.
[134]Van Hoof D, PassierR,Ward-Van Oostwaard D,et al.A quest for human andmouse embryonic stem cell specific proteins[J].Mol Cell Proteomics, 2006,5(7):1261-1273.
[135]Schulz TC, Swistowska AM,Liu Y,etal.A large-scale proteomic analysis of human embryonic stem cells[J]. BMC Genomics, 2007,8: 478.
[136]Seshi B. An integrated approach to mapping the proteome of the human bone marrow stromal cell[J]. Proteomics,2006,6(19):5169-5182.
[137]Foster LJ, Zeemann PA,LiC,et al.Differential expression profiling Of membrane proteins by quantitative proteomics in a human mesenchymal stem cell line undergoing osteoblast differentiation[J].Stem Cells,2005, 23(9):1367-1377.
[138]刘元林,于晓妉,刘晓丹,等.间充质干细胞定向成骨分化的蛋白质组学分析[J].第四军医大学学报, 2006, 27 (17 ):1551-1554.
[139]Mito Toyoda , Yoshinori Matsubara , Konghua Lin. Characterization and comparison of adipose tissue-derived cells from human subcutaneous and omental adipose tissues. Cell Biochem Funct[J].2009,27:44-447.
[140]Lange C, Schroeder J, Stute N,et al. High-potential human mesenchymal stem cells[J].Stem Cells Dev,2005,14:70-80.
[141] Akino K, Mineta T, Fukui M,et al.Bone morphogenetic protein-2 regulates proliferation of human mesenchymal stem cells[J]. WOUND REPREG,2003,11:354-360.
[142]成令忠.组织学.2版.人民卫生出版社,1993:82-87.
[143]Bordji K, Grillasca JP, Gouze JN,et al. Evidence for the presence of peroxisome proliferator-activated receptor (PPAR) alpha and gamma and retinoid Z receptor in cartilage.PPARgamma activation modulates the effects of interleukin-1beta on rat chondrocytes [J]. J Biol Chem,2000, 275(16):12243-12250.
[144]Shu-Ching Shih and Kevin P. Claffey. Regulation of Human Vascular Endothelial Growth Factor mRNA Stability in Hypoxia by Heterogeneous Nuclear Ribonucleoprotein L[J].BIOLOGICAL CHEMISTRY,1999,274(3): 1359–1365.
[145]Julie S. Lau,Peter Baumeister,Edmund Kim, et al. Heterogeneous Nuclear Ribonucleoproteins as Regulators of Gene Expression Through Interactions With the Human Thymidine Kinase Promoter.Cellular Biochemistry[J]. 2000, 79:395–406.
[146]Denis Feliers,Myung-Ja Lee,Goutam Ghosh-Choudhury,et al. Hetero- geneous nuclear ribonucleoprotein K contributes to angiotensin II stimul- ation of vascular endothelial growth factor mRNA translation[J]. Am J Physiol Renal Physiol,2007, 293: F607–F615.
[147] James R. Stone, Tucker Collins. Rapid Phosphorylation of Heterogeneous Nuclear Ribonucleoprotein C1/C2 in Response to Physiologic Levels of Hydrogen Peroxide in Human Endothelial Cells[J].BIOLOGICAL CHEM- ISTRY,2002,277(18): 15621–15628.
[148] Mechthild H,Frank WW.Formation In Vitro of Heterotypic Complexes and Intermediate-sized Filaments by Homologous and Heterologous Recombinations of Purified Polypeptides[J].CELL BIOLOGY, 1985,101:1826-1841.
[149]Filion M, Sarafian V, Lussier M,et al. Arrangement of a Cluster of ThreeMouse Type I Keratin Genes Expressed Sequentially during Esophageal- Type Epithelial Cell Differentiation. Genomics[J]. 1994,24(2):303-10.
[150]Chuang N.N,Huang C.C.Interaction of integrin beta1 with cytokeratin 1 in neuroblastoma NMB7 cells.Biochem Soc Trans[J]. 2007,35:1292-1294.
[151]Spichkina OG, Pinaev GP, Petrov IuP. Analysis of heterogeneity of human keratinocyte populations by their adhesion to substrate and the keratin 19 to actin ratio. Tsitologiia[J].2008,50(3):218-27.
[152]Blachly-Dyson E, Zambronicz EB, Yu WH, et al.Cloning and functional expression in yeast of two human isoforms of the outer mitochondrial membrane channel, the voltage-dependent anion channel[J].J Biol Chem. 1993,268(3):1835-1841.
[153]陆秋涯,黄立东,陆佩华. Annexin A2的结构和功能及其与疾病的关系.医学分子生物学杂志[J]. 2008, 5 (2) : 1742 176.
[2]NIH. Stem Cells : Scientific Progress and Future Research Directions.(August 6 ,2001) Website : http :/ / www. nih. g ov/ news/ stemcell/ scireport . htm
[3]Hochedlinger K,Jaenisch R.Nuclear reprogramming and pluripotency [J]. Nature, 2006,441(7097):1061-1067.
[4]Rodolfa KT, Eggan K.A transcriptional logic for nuclear reprogramming [J].Cell,2006,126(4):652-655.
[5]Cervera RP,Stojkovic M.Commentary:somatic cell nuclear transfer-progress and promise[J].Stem Cells,2008,26(2):94- 495.
[6]Daley GQ, Goodell MA, Snyder EY. Realistic prospects for stem cell therapeutics[R].Hematology Am Soc Hematol Educ Program,2003: 398-418.
[7]Evans M J, Kaufman M H. Establishment in culture of pluripotent cells from mouse embryos[J]. Nature,1981,292(9):154-156.
[8]Takahashi K,Yamanaka S.Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors[J]. Cell, 2006, 126(4):663-676.
[9]Hyun I,Hochedlinger K, Jaenisch R,et al. New advances in iPS cell research do not obviate the need for human embryonic stem cells[J].Cell Stem Cell, 2007,1(4): 367-368.
[10]Watts G. Scientists“reprogramme”skin cells to create embryonic stem cells[J]. BMJ, 2007,335(7630): 1112-1113.
[11]Cyzy J, Wiese C, Rolletschek A,et al.Potential of embryonic and adult stem cells in vitro[J]. Biol Chem,2003,384(10-11):1391-1409.
[12]程冬婉,张伟.成体干细胞临床应用的研究[J].中国肿瘤生物治疗杂志,2006,13(2):150-152.
[13]周长梅.干细胞研究进展[J].淮北煤炭师范学院学报,2006,27(4):45-48.
[14]Watts C, McConkey H, Anderson L,et al.Anatomical perspectives onadultneural stem cells[J]. JAnat, 2005, 207(3): 197-208.
[15]SchafflerA, BuchlerC.Concise review: adipose tissue-derived stromal cells--basic and clinical implications fornovel cell-based therapies[J]. Stem Cells, 2007,25(4): 818-827.
[16]Mimeault M, Hauke R,Mehta PP,etal.Recent advances in cancer stem /progenitor cell research: therapeutic implications for overcoming resistance to the most aggressive cancers[ J]. J Mol Cell Med, 2007, 11(5): 981-1011.
[17]Minguell JJ, Er ices A,Conget P. Mesenchymal stem cells[J]. Exp Biol Med, 2001, 226 (6) : 507-520.
[18]曹谊林,崔磊,刘伟.组织工程学理论与实践[M].北京:科学出版社,2008年,第一版.
[19]Ng AM, Kojima K, Kodoma S, et al. Isolation techniques of murine bone marrow progenitor cells and their adipogenic, neurogenic and osteogenic differentiation capacity[J]. Med J Malaysia. 2008,63:121-122.
[20]Zuk PA, Zhu M, Ashjian P, et al.Human adipose tissue is a source of multipotent stem cells[J]. Mol Biol Cell 2002,13: 4279–4295
[21]Gronthos S, Franklin DM, Leddy HA, et al. Surface proteincharacterization of human adipose tissue- derived stromal cells [J]. J Cell Physiol, 2001, 189: 54- 63.
[22]Safford KM, Hicok KC, Safford SD, et al. Neurogenic differentiation of urine and human adipose- derived stromal cells [J]. Biochem Biophys Res Commun,2002,294:371- 379.
[23]Miranville A, Heeschen C, Sengenes C, et al. Improvement of postnatal neovascularization by human adipose tissue- derived stem cells [J]. Circulation, 2004,20:349- 355.
[24]Planat- Benard V, Silvestre JS, Cousin B, et al. Plasticity of human adipose lineage cells toward endothelial cells:physiological and therapeutic perspec- tives[J]. Circulation,2004,10:656- 663.
[25]De Ugarte DA, Morizono K, Elbarbary A, et al.Comparison of multi-lineage cells from human adipose tissue and bone marrow[J]. Cells Tissues Organs 2003,174: 101-109.
[26]Izadpanah R, Trygg C, Patel B, et al. Biologic properties of mesenchymal stem cells derived from bone marrow and adipose tissue[J].J Cell Biochem, 2006,99:1285-1297.
[27]Zuk PA, Zhu M, Mizuno H, et al. Multilineage cells from human adipose tissue: implications for cell-based therapies[J].Tissue Eng 2001,7: 211–228.
[28]Mizuno H, Hyakusoku H. Mes engenic potential and future clinical perspective of human proces sed lipoaspirate cel ls [J]. J Nippon Med Sch, 2003,70(4):300- 306.
[29]金杰,虞渝生.脂肪干细胞研究进展.浙江医学.2008,30(1):101-103.
[30]Lee RH,Kim B,Choi I,etal.Characterization and expression analysis ofmesenchymal stem cells from human bonemarrow and adipose tissue [J]. Cell Physiol Biochem, 2004,14(4/6): 311-324.
[31]Gimble J, Guilak F. Adipose-derived adult stem cells: isolation, characteri- zation,and differentiation potential [J]. Cytotherapy, 2003,5(5): 362-369.
[32] Guilak F, Lott KE, Awad HA, et al. Clonal Analysis of the Differentiation Potential of Human Adipose-Derived Adult Stem Cells[J].J Cell Physiol, 2006,206: 229–237.
[33]Mi Hyang Moon, Sun Young Kim, Yeon Jeong Kim1,et al. Human Adipose Tissue-Derived Mesenchymal Stem Cells Improve Postnatal Neovascula- rization in a Mouse Model of Hind limb Ischemia[J]. Cell Physiol Biochem, 2006,17:279-290.
[34]Ying Cao, Zhao Sun, Lianming Liao,et al. Human adipose tissue-derived stem cells differentiate into endothelial cells in vitro and improve postnatal neovascularization in vivo[J]. Biochemical and Biophysical Research Communications,2005,332:370–379.
[35]E. Raposio, C. Guida, I. Baldelli,et al. Characterization and induction ofhuman pre-adipocytes[J]. Toxicology in Vitro,2007, 21:330–334.
[36]Naoki Yamamoto, Hirohiko Akamatsu, Seiji Hasegawa,et al. Isolation of multipotent stem cells from mouse adipose tissue[J]. Journal of Dermat- ological Science,2007,48:43-52.
[37]De Ugarte DA, Alfonso Z, Zuk PA,et al. Differential expression of stem cell mobilization-associated molecules on multi-lineage cells from adipose tissue and bone marrow[J].Immunol Lett,2003,89:267–270.
[38]Lin TM, Tsai JL, Lin SD,et al. Accelerated growth and prolonged lifespan of adipose tissue-derived human mesenchymal stem cells in a medium using reduced calcium and antioxidants[J].Stem Cells Dev, 2005,14(1):92–102.
[39]Tholpady SS, Llull R, Ogle RC,et al.Adipose tissue: stem cells and beyond [J].Clin Plast Surg,2006,33(1):55–62.
[40]Gimble JM. Adipose tissue-derived therapeutics[J].Expert Opin Biol Ther, 2003,3(5):705–13.
[41]Cheryl T. Gomillion, Karen J.L. Burg. Stem cells and adipose tissue engineering[J].Biomaterials,2006,27: 6052–6063.
[42]Katz A J, Ashok T, Sunil S T,et al. Cell surface and transcriptional characterization of human adipose-derived adherent stromal(hADAS) cells [J]. Stem Cells,2005,23:412-423.
[43]Strem BM, Hicok KC, Zhu M,et al. Multipotential differentiation of adipose tissue-derived stem cells[J]. Keio J Med,2005,54(3):132-141.
[44]郭新,赵东海,何旭等.大鼠骨髓间充质干细胞的分离培养[J].吉林大学学报:医学版,2005,31(3):469-471.
[45]崔磊,尹烁,杨平等.脂肪干细胞HLA分子表达与体外抑制淋巴细胞增殖的实验研究[J].中华医学杂志,2005,85(27):1890-1894.
[46]Aust L, Devlin B, Foster SJ,et al.Yield of human adiposederived adult stem cells from liposuction aspirates[J].Cytotherapy, 2004,6:7-14.
[47]Lee RH, Kim B, Choi I,et al.Characterization and expression analysis ofmesenchymal stem cells from human bone marrow and adipose tissue[J].Cell Physiol Biochem,2004,14:311-324.
[48]Planat-Benard V, Silvestre JS, Cousin B,et al. Plasticity of human adipose lineage cells toward endothelial cells: physiological and therapeutic perspectives[J].Circulation,2004,109:656–663.
[49]Rangappa S, Fen C, Lee EH,et al.Transformation of adult mesenchymal stem cells isolated from the fatty tissue into cardiomyocytes[J]. Ann Thorac Surg,2003,75:775-779.
[50]Safford KM, Hicok KC, Safford SD,et al.Neurogenic differentiation of murine and human adipose-derived stromal cells[J].Biochem Biophys Res Commun,2002,294:371-379.
[51]Ying Cao, Zhao Sun, Lianming Liao,et al. Human adipose tissue -derived stem cells differentiate into endothelial cells in vitro and improve postnatal neovascularization in vivo[J]. Biochemical and Biophysical Research Communications,2005,332:370-379.
[52]Strem BM, Hicok KC, Zhu M,et al. Multipotential differentiation of adipose tissue-derived stem cells[J]. Keio J Med, 2005,54:132-141.
[53]Slaek,JM. Stem ells in Epithelial Tissues[J].Science,2000,287: 1431-1433.
[54]He X, Li YL, Wang XR,et al. Mesenchymal stem cells transduced by PLEGFP-N1 retroviral Vector maintain their biological features and differentiation[J].Chin Med J,2005,118:1728-1734.
[55]李慧武,戴赶戎,汤亭亭等.长期体外培养对人脂肪源性间充质干细胞生物学特性的影响[J].中华创伤骨科杂志,2006,8(10):929-932.
[56]Bartholomew A,Sturgeon C,Siatskas M,et al.Mesenehymal stem cells suppress lymphocyte Proliferation in vitro and Prolong skin graft survival in vivo[J].Exp Hematol,2002,30:42-48.
[57]Ogawa R, Mizuno H, Hyakusoku H,et al. Chondrogenic and osteogenic differentiation of adipose derived stem cells isolated from GFP transgenicmice[J]. J Nippon Med Sch,2004,71(4):240-241.
[58]Huang J I, Zuk P A, Jones N F,et al. Chondrogenic potential of multipotential cells from human adipose tissue[J]. Plast Recons trSurg,2004, 113(2): 585- 594.
[59]杨亚军,朱庆生. CDMPI诱导大鼠脂肪干细胞体外成软骨细胞的实验[J].第四军医大学学报,2008,29(4):342-345.
[60]陈崎,彭智.脂肪干细胞在骨、软骨组织工程中的研究进展[J].中国民族民间医药.2009,14:26-27.
[61]Liu Y, LuoE, Chen X,et al. Molecular and cellular characterization during chondrogenic differentiation of adipose tissue-derived stromal cells in vitro and cartilage formarionin vivo[J]. J Cell Mol Med,2005,9(4):929-939.
[62]Mochizuki T, Muneta T, SakaguchiY,et al. Higher chondrogenic potentialof fibrous synovium-and adipose synovium-derived cells compared with subcutaneous fat-derived cells:Distinguishingproperties ofmesenchymal stem cells in humans[J]. ArthritisRheum,2006,54(3):843-853.
[63]SegawaY, MunetaT, MakinoH,et al. Mesenchymal stem cells derived from synovium, meniscus, anterior cruciate ligament,and articular chondrocytes share similar gene expression profiles[J]. J Orthop Res,2009,27(4):435-441.
[64]Huang J I, Zuk P A, Jones N F, et al. Chondrogenic potential of multipotential cells from human adipose tissue[J]. Plast Recons trSurg, 2004, 113(2):585-594.
[65]杨平,尹烁,崔磊等.脂肪干细胞向血管平滑肌细胞诱导的实验研究[J].中国修复重建外科杂志,2008,22:481-486.
[66]张端珍,盖鲁粤,刘宏伟等.脂肪干细胞诱导成心肌细胞研究[J].心脏杂志,2005,17:405-408.
[67]Mizuno H, Zuk PA, Zhu M,et al. Myogenic differentiation by human processed lipoaspirate cells[J]. Plast Reconstr Surg,2002,109(1): 199-209.
[68]何旭,王心蕊,张海英等.人骨髓间充质干细胞体外定向诱导分化为脂肪细胞方法的建立[J].吉林大学学报,2006,3(32):530-532.
[69]Herrera-Herrera M, Zapata-Bustos R, Salazar-Olivo L. Simplified culture techniques for growth and differentiation of murine and human pre-adipocytes for translational applications[J]. Cytotherapy, 2009, 11(1): 52-60.
[70]张勇.脂肪干细胞的研究进展[J].医学研究生学报,2009,22(10):1088- 1092.
[71]Park JP, Jung JW, Lee YS,et al. The roles of Wnt antagonists Dkk1 and sFRP4 during adipogenesis of human adipose tissue-derived mesenchymal stem cells[J]. Cell Prolif,2008, 41(6):859-874.
[72]Scheideler M, Elabd C, Zaragosi LE,et al.Comparative transcriptomics of human multipotent stem cells during adipogenesis and osteoblastogenesis [J].BMC Genomics,2008, 17(9):340.
[73]Chen J, Zhang ZG, Ly Y,et al.Intravenousadministration of human bone marrow stromal cells induces angiogenesis in the ischemic boundary zone after stroke in rats[J].Circ Res,2003,92(6):692-699.
[74]Haynes worth SE, Baber MA, Caplan AI. Cell surface antigens on human marr ow-derived mesenchymal cells are detected by monoclonal antibodies [J]. Bone,1992,13:68-69.
[75]Barry FP,Boynton RE,Haynesworth S,et al.The monoclonal antibody SH-2,raised against human mesenchymal stem cells,recognizes an epitope on endoglin(CD105)[J].Biochem Biophys Res Commun, 1999,265(1):134-139.
[76]Fleming JE,Haynesworth SE,Cassiede P,et al.Monoclonal antibody against adult marrow-derived mesenchymal stem cells recognizes developing vasculature in embryonic human skin[J].Dev Dyn,1998,212(1):119-132.
[77]Ohneda O,Ohneda K,Arai F,et al.ALCAM(CD166):its role in hematopoietic and endothelial development[J].Blood,2001,98 (7):2134-2142.
[78]Reyes M,Verfaillie CM.Characterization of multipotent adult progenitor cells,a subpopulation of mesenchymal stem cells[J].Ann NY Acad Sci, 2001,938:231-235.
[79]Reyes M,Dudek A,Jahagirdar B,et al.Origin of endothelial progenitors in human postnatal bone marrow[J].J Clin Invest,2002,109(3):337-346.
[80]Lozito TP, Kuo CK, Taboas JM,et al. Human Mesenchymal Stem Cells Express Vascular Cell Phenotypes Upon Interaction With Endothelial Cell Matrix[J]. Cellular Biochemistry,2009,107:714–722.
[81] Oswald J, Boxberger S, J?rgensen B,et al. Mesenchymal Stem Cells Can Be Differentiated Into Endothelial Cells In Vitro[J].STEM CELLS.2004,22: 377- 384.
[82]Chen MY, Lie PC, Li ZL,et al. Endothelial differentiation of Wharton's jelly–derived mesenchymal stem cells in comparison with bone marrow–derived mesenchymal stem cells[J].Exp Hematol,2009,37: 629-640.
[83]Valérie Planat-Benard, Jean-Sébastien Silvestre, Béatrice Cousin,et al. Plasticity of Human Adipose Lineage Cells Toward Endothelial Cells Physiological and Therapeutic Perspectives[J].Circulation,2004,109:656- 663.
[84] De Francesco F, Tirino V, Desiderio V,et al. Human CD34+/CD90+ASCs Are Capable of Growing as Sphere Clusters, Producing High Levels of VEGF and Forming Capillaries[J].PLoS ONE,2009,4(8):1-13.
[85]Ying Cao, Zhao Sun, Lianming Liao,et al.Human adipose tissue-derived stem cells differentiate into endothelial cells in vitro and improve postnatal neovascularization in vivo[J]. Biochemical and Biophysical Research Communications,2005,332: 370-379.
[86]Watt F,Hogan B.Out of Eden:stem cells and their niches[J]. Science,2000, 287(5457):1427-1430.
[87]Roberts DM,Kearney JB,Johnson JH,et al.The vascular endothelial growth factor(VEGF) receptor Flt-1(VEGFR-1) modulates Flk-1(VEGFR-2) signaling during blood vessel formation[J].Am J Pathol,2004, 164(5): 1531- 1535.
[88]Kearney JB,Kappas NC,Ellerstrom C,et al.The VEGF receptor flt-1 (VEGFR-1) is a positive modulator of vascular sprout formation and branching morphogenesis[J].Blood,2004,103(12):4527-4535.
[89]Wang M,Crisostomo PR,Herring C,et al.Human progenitor cells from bone marrow or adipose tissue produce VEGF,HGF,and IGF-I in response to TNF by a p38 MAPK-dependent mechanism[J].Am J Physiol Regul Integr Comp Physiol,2006,291(4):880-884.
[90]Zeng Q,Li X,Beck G,et al.Growth and differentiation factor-5(GDF-5) stimulates osteogenic differentiation and increases vascular endothelial growth factor(VEGF) levels in fat derived stromal cells in vitro[J].Bone, 2007,40(2):374-381.
[91]Crisostomo PR,Wang M,Herring CM,et al.Gender differences in injury induced mesenchymal stem cell apoptosis and VEGF,TNF,IL-6 expression: Role of the 55 kDa TNF receptor(TNFR1)[J].J Mol Cell Cardiol, 2007,42(1):142-149.
[92]Oswald J,Boxberger S,Jorgensen B,et al.Mesenchymal stem cells can be differentiated into endothelial cells in vitro[J].Stem Cells,2004,22(3): 377- 384.
[93]Al-Khaldi A,Eliopoulos N,Martineau D,et al.Postnatal bone marrow stromal cells elicit a potent VEGF-dependent neoangiogenic response in vivo[J].Gene Ther,2003,10(8):621-629.
[94]Annabi B,Naud E,Lee YT,et al.Vascular progenitors derived from murine bone marrow stromal cells are regulated by fibroblast growth factor and are avidly recruited by vascularizing tumors[J].J Cell Biochem,2004,91(6): 1146-1158.
[95]Adams JC,Watt FM. Regulation of development and differentiation by the extra cellular matrix[J].Development,1993,117(4):1183- 1198.
[96]郭新,李伟,朱桂彬等.基质蛋白与整合素参与人骨髓间充质干细胞体外内皮的分化[J].中国临床康复,2005,9(14):58-60.
[97]Meredith JE,Fazeli B,Schwartz MA.The extracellular matrix as a cell survival factor[J].Mol Biol Cell,1993,4(9):953-961.
[98]Annabi B,Lee YT,Turcotte S,et al. Hypoxia promotes murine bone- marrow- derived stromal cell migration and tube formation[J].Stem Cells,2003,21 (3):337-347.
[99]Remy-Zolghadri M. Endothelium ProPerties of a tissue-engineered blood Vessel for small-diameter vascular reconstruetion[J].J Vase Surs,2004,39(3): 613-620.
[100]Dejana,E.Endothelial cell-cell junctions:happy together[J].Nat Rev Mol Cell Biol,2004,5(4):261-270.
[101]Feraud,0,Y.Cao,D.Vittet.Embryonic stem cell-derived embryoid bodies development in collagen gels recapitulates sprouting angiogenesis[J].Lab Invest,2001,81(12):1669-1681.
[102]Feraud,0.,M.H.Prandini,D.Vittet,Vaseulogenesis and angiogenesis from in vitro differentiation of mouse embryonic stem cells[J]. Methods Enzymol, 2003,365:214-228.
[103]Mallet,C.TGFbetal induces vaseulogenesis and inhibits angiogenic Sprout- ing in an embryonic stem cell differentiation model: respeetive Contribution of ALK1 and ALK5[J].Stem Cells.2006, 24(11):2420-2427.
[104]Lyden,D.Impaired recruitment of bone-marrow-derived endothelial and he- matopoietic Precursor cells blocks tumor angiogenesis and growth[J].Nat Med,2001,7(11):1194-1201.
[105]Wikins MR,Pasquali C.From Proteins to Proteomes:Large scalc protein identification by two-dimensional electrophoresis and amino acid analysis [J].Bio/Technology,1996,14:6561-6567.
[106]Petricoin EF,Zoon KC.Clinical proteomics:translating benchside promise into bedside reality[J].Nature Rev Drug Discov,2002, 1(9):683-695.
[107]Petricoin EF,Zoon KC.Clinical proteomics:translating benchside promise into bedside reality[J].Nature Rev Drug Discov,2002,1(9):683-695.
[108]P.Kahn,From genome to Proteome:looking at a cell proteins[J]. Science, 1995,270:369-370.
[109]D.Swinbanks,Govenmment backs proteome proposal[J].Nature, 1995,378: 653.
[110]Beranova G,Iorgianni S.Proteome analysis by two-dismensional gel electrophoresis and mass spectrometry:strengths and limitations [J].Trends in Analytical Chemistry,2003,22(5): 273-280.
[111]O Parrell PH.High resolution two-dimensional elcetrophoresis of proteins [J].Journal of Biological Chemistry,1975,250:4007 -4021.
[112]Li C,Tan YX,Zhou H,et al. Proteomic analysis of hepatitis B virus- associated hepatocellular carcinoma:Identification of Potential tumor markers[J].Proteomics,2005,5(4):1125-1139.
[113]Sun W,Xing B,Sun Y,et al. Proteome analysis of hepatocellular carcinoma by two-dimensional Difference gel electrophoresis: novel protein markers in hepatocellular carcinoma tissues[J]. MOL Cell Proteomics,2007,6(10): 1798-1808.
[114]Blanc JF,Lalanne C,Plomion C,et al.Proteomic analysis of Differentially expressed Proteins in hepatocellular carcinoma developed in Patients with chronic viral hepatitis C[J]. Proteomics,2005,5(14):3778-3789.
[115]Kang SM,Shin MJ,Kim JH,et al.Proteomic Profiling of cellular Proteins Interacting with the hepatitis C virus core Protein[J]. Proteomics,2005,5(8): 2227-2237.
[116]Liang CR,Leow CK,Neo JC,et al.Proteome analysis of human hepatocell ular carcinoma tissues by two-dimensional difference gel electrophoresis and mass spectrometry[J]. Proteomics,2005, 5(8):2258-2271.
[117]Jorgenson JW,Lukacs KD.Zone electrophoresis in open-tubular glasscapillareies[J].Anal Chem,1981,53:1298.
[118]Garcia Campana AM,Gamiz Gracia L,Baeyens WR,et al.Dervatization of bicmolecules for chemilum inescent detection in capillary electrophor esis[J]. Chromatogr B Analyt Technol Bicmed Life Sci,2003,793:49-74.
[119]Shevchenko A,Loboda A,Shevchenko A,et al. MALDI quadrupole time-of– flight mass spectrometry:a powerful tool for proteomic research[J].Anal Chem,2000,72:2132-2141.
[120]Minden J,Waggoner A.[R].US patent:6043025,2000.
[121]Venter JC,Adams MD,Myers EW,et al.The sequence of the humangen- ome[J].Science,2001,291(5507):1304-1351.
[122]Mike Tyers,Matthias Mann M.From genomics to Proteomics[J]. Nature, 2003,422(6928):193-197.
[123]Pandey A,Manll M.Proteomics to study genes and genomes[J].Nature, 2000,405(6788):837-846.
[124]Ruedi Aebersold,Matthias Mann. Mass spectrometry-based Proteomics [J]. Nature,2003,422(6928):198-207.
[125]Ong SE,Mann M.Mass spectrometry-based Proteomics turns quantitative [J].Nature,Chemical Biology,2005,l(5):252-262.
[126]Zvonic S,Lefevre M, Kilroy G,et al.Secretome of primary cultures of human adipose-derived stem cells:modulation of serpins by adipogenesis[J].Mol Cell Proteomics,2007,6 (1):18-28.
[127]Pittenger MF, Mackay AM, Beck SC,et al. Multilineage potential of adult human mesenchymal stem cells[J]. Science.1999, 284: 143–147.
[128]李文婷,孙华林,许增禄等。大鼠骨髓基质细胞向Schwann细胞分化后蛋白表达变化的研究[J].中国科学,2009,39(2):167-178.
[129]Wang D,Gao L.Proteomic analysis of neural differentiation of mouse embryonic stem cells[J].Proteomics,2005,5(17):4414- 4426.
[130]Elliott ST,Crider DG, Garnham CP,et al. Two-dimensional gel electrop-horesis database of murine R1 embryonic stem cells[J].Proteomics, 2004,4(12):3813-3832.
[131]Wang J,RaoS,Chu J,etal.A protein interaction network for pluripotency of embryonic stem cells[J].Nature,2006,444(7117): 364-368.
[132]Prudhomme W,Daley GD, Zandstra P,etal.Multivariate proteomic analysis ofmurine embryonic stem cel self-renewal versus differentiaion signaling[J]. ProcNatlAcad SciUSA, 2004, 101(9): 2900-2905.
[133]Baharvand H, Fathi A, Gourabi H,et al. Identification of mouse embryonic stem cell-associated proteins[J]. JProteomeRes, 2008,7(1): 412-423.
[134]Van Hoof D, PassierR,Ward-Van Oostwaard D,et al.A quest for human andmouse embryonic stem cell specific proteins[J].Mol Cell Proteomics, 2006,5(7):1261-1273.
[135]Schulz TC, Swistowska AM,Liu Y,etal.A large-scale proteomic analysis of human embryonic stem cells[J]. BMC Genomics, 2007,8: 478.
[136]Seshi B. An integrated approach to mapping the proteome of the human bone marrow stromal cell[J]. Proteomics,2006,6(19):5169-5182.
[137]Foster LJ, Zeemann PA,LiC,et al.Differential expression profiling Of membrane proteins by quantitative proteomics in a human mesenchymal stem cell line undergoing osteoblast differentiation[J].Stem Cells,2005, 23(9):1367-1377.
[138]刘元林,于晓妉,刘晓丹,等.间充质干细胞定向成骨分化的蛋白质组学分析[J].第四军医大学学报, 2006, 27 (17 ):1551-1554.
[139]Mito Toyoda , Yoshinori Matsubara , Konghua Lin. Characterization and comparison of adipose tissue-derived cells from human subcutaneous and omental adipose tissues. Cell Biochem Funct[J].2009,27:44-447.
[140]Lange C, Schroeder J, Stute N,et al. High-potential human mesenchymal stem cells[J].Stem Cells Dev,2005,14:70-80.
[141] Akino K, Mineta T, Fukui M,et al.Bone morphogenetic protein-2 regulates proliferation of human mesenchymal stem cells[J]. WOUND REPREG,2003,11:354-360.
[142]成令忠.组织学.2版.人民卫生出版社,1993:82-87.
[143]Bordji K, Grillasca JP, Gouze JN,et al. Evidence for the presence of peroxisome proliferator-activated receptor (PPAR) alpha and gamma and retinoid Z receptor in cartilage.PPARgamma activation modulates the effects of interleukin-1beta on rat chondrocytes [J]. J Biol Chem,2000, 275(16):12243-12250.
[144]Shu-Ching Shih and Kevin P. Claffey. Regulation of Human Vascular Endothelial Growth Factor mRNA Stability in Hypoxia by Heterogeneous Nuclear Ribonucleoprotein L[J].BIOLOGICAL CHEMISTRY,1999,274(3): 1359–1365.
[145]Julie S. Lau,Peter Baumeister,Edmund Kim, et al. Heterogeneous Nuclear Ribonucleoproteins as Regulators of Gene Expression Through Interactions With the Human Thymidine Kinase Promoter.Cellular Biochemistry[J]. 2000, 79:395–406.
[146]Denis Feliers,Myung-Ja Lee,Goutam Ghosh-Choudhury,et al. Hetero- geneous nuclear ribonucleoprotein K contributes to angiotensin II stimul- ation of vascular endothelial growth factor mRNA translation[J]. Am J Physiol Renal Physiol,2007, 293: F607–F615.
[147] James R. Stone, Tucker Collins. Rapid Phosphorylation of Heterogeneous Nuclear Ribonucleoprotein C1/C2 in Response to Physiologic Levels of Hydrogen Peroxide in Human Endothelial Cells[J].BIOLOGICAL CHEM- ISTRY,2002,277(18): 15621–15628.
[148] Mechthild H,Frank WW.Formation In Vitro of Heterotypic Complexes and Intermediate-sized Filaments by Homologous and Heterologous Recombinations of Purified Polypeptides[J].CELL BIOLOGY, 1985,101:1826-1841.
[149]Filion M, Sarafian V, Lussier M,et al. Arrangement of a Cluster of ThreeMouse Type I Keratin Genes Expressed Sequentially during Esophageal- Type Epithelial Cell Differentiation. Genomics[J]. 1994,24(2):303-10.
[150]Chuang N.N,Huang C.C.Interaction of integrin beta1 with cytokeratin 1 in neuroblastoma NMB7 cells.Biochem Soc Trans[J]. 2007,35:1292-1294.
[151]Spichkina OG, Pinaev GP, Petrov IuP. Analysis of heterogeneity of human keratinocyte populations by their adhesion to substrate and the keratin 19 to actin ratio. Tsitologiia[J].2008,50(3):218-27.
[152]Blachly-Dyson E, Zambronicz EB, Yu WH, et al.Cloning and functional expression in yeast of two human isoforms of the outer mitochondrial membrane channel, the voltage-dependent anion channel[J].J Biol Chem. 1993,268(3):1835-1841.
[153]陆秋涯,黄立东,陆佩华. Annexin A2的结构和功能及其与疾病的关系.医学分子生物学杂志[J]. 2008, 5 (2) : 1742 176.