脐带间充质干细胞对CD34~+祖细胞免疫反应的影响
|
摘要
目的探讨缺氧及非缺氧脐带来源的间充质干细胞(UC-MSCs)是否能有效抑制脐血CD34+祖细胞免疫反应,影响其反应过程中免疫细胞因子的分泌。
方法通过胶原酶及胰酶消化法从脐带中分离获得MSCs,观察其生长形态,取传代培养至第3代的UC-MSCs流式细胞术检测其表面标志；淋巴细胞分离液分离外周血单个核细胞(PBMC);免疫磁珠分离脐血CD34+祖细胞；将不同数量的第三代UC-MSCs5×103、2×104、8×104分别加入脐血CD34+祖细胞、外周血单个核细胞共培养体系中,不加UC-MSCs的CD34+祖细胞、PBMC共培养体系为阴性对照；另设用Transwel1悬挂式小室将UC-MSCs和脐血CD34+祖细胞、PBMC共培养体系分隔培养；培养96 h后,分别收集上清液,ELISA检测IFN-y和IL-10分泌水平。氯化钴(COCl2)进行化学模拟缺氧,在UC-MSCs中加入氯化钴浓度为0、50、100、150、200、250μmol/L的细胞培养液,分别于12 h、1d、2 d、3d、4d、5d、6 d通过MTT法检测各组的OD值,从而确定各组细胞的增殖情况；将氯化钻浓度为150μmol/L的培养液缺氧处理脐带MSCs48h后,加到脐血CD34+祖细胞、PBMC共培养体系中,培养96 h后,收集上清液,ELISA检测IFN-y和IL-10分泌水平。
结果脐带UC-MSCs高表达CD 105、CD90、CD29、CD49,不表达CD40、CD80、CD86、HLA-DR、CD34、CD45;加UC-MSCs组IFN-γ的分泌量减少(与阴性对照组比,P<0.05),IL-10的分泌量增多(与阴性对照组比,P<0.05),而且具有数量依赖性,即数量越多免疫抑制作用越强；Transwell分隔培养组IFN-γ的分泌量减少(与阴性对照组相比,P<0.05),IL-10的分泌量增多(与阴性对照组相比,P<0.05)；但与接触培养组相比,IFN-y的分泌量增多,IL-10的分泌量减少。经氯化钴模拟缺氧处理的UC-MSCs(150μmol/L)其对数生长期提前,对数期缩短,增殖速度加快,第3天之后减慢,提前达平台期,4-5 d后呈逐渐降低趋势,而经较高浓度(200、250μmol/L)CoCl2处理的UC-MSCs生长曲线趋于低平。缺氧的脐带MSCs能抑制共培养体系中淋巴细胞IFN-y的分泌(与阴性对照组比,P<0.05),同时促进IL-10分泌(与阴性对照组比,P<0.05),但与非缺氧脐带MSCs组相比,IFN-y的分泌量增多,IL-10的分泌量减少。
结论通过酶消化法可从脐带中成功分离获得MSCs;氯化钴(浓度≤150μmol/L)对UC-MSCs增殖影响微弱,可用于其模拟缺氧预处理；缺氧／非缺氧UC-MSCs都能抑制脐血CD34+祖细胞的免疫反应中IFN-γ的分泌,同时促进IL-10分泌,并且这种作用部分依赖于细胞间的相互接触；缺氧处理后的UC-MSCs免疫调节功能下调；UC-MSCs与脐血CD34+祖细胞联合移植可有效减轻同种异体干细胞移植后的免疫排斥反应,提高干细胞的生存率,为治疗缺血性心肌病提供有效的方法。
Objective To investigate wheather the hypoxic/non-hypoxic umbilical cord-derived mesenchymal stem cells can inhibit the immune responses of umbilical cord blood (UCB)CD34+ progenitor cells, affecting the secretion of immunomodulattory cytokines.
Methods UC-MSCs were isolated and cultured, the growth morphology were observed. Surface markers of UC-MSCs were detected by flow cytometry. The CD34 +progenitor cells in umbilical blood were isolated by immunomagnetic beads. Peripheral blood mononuclear cells (PBMCs) were isolated from healthy volunteers using Ficoll-Histopaque density gradients from heparinized samples.The different number of UC-MSCs(5×103、2×104、8×104)were added to the co-culture system of UCB CD34+ progenitor cells and peripheral blood mononuclear cells(PBMC),and cultivation of CD34+ cells and PBMC as negative group, in addition, we mixed this three kinds of cells using Transwell,UBC CD34+ cells and PBMC were cultured in the upper separated from UC-MSCs, after culturing for 96 h, supernatants were collected and detected of the IFN-y and IL-10 level by ELISA. Third passage (P3) UC-MSCs were separately cultivated in DMEM/F12 with cobalt chloride 0、50、100、150、200、250μmol/L at 37℃and 5% CO2 for 12h、1d、2d、3d、4d、5d、6d, detected the proliferation and vitality of every groups by MTT assay. The hypoxic UC-MSCs which were cultivated with CoCl2150μmol/L for 48h were added to the co-culture system of UBC CD34+ cells and PBMC, culturing for 96 h, detected cytokines using above method.
Results UC-MSCs did not express CD40, CD80, CD86, HLA-DR, CD34.It can inhibit IFN-y secretion (compare with controls,P<0.05), promote IL-10 secretion(compared with controls,P<0.05) and had a dose-depended. Using Transwell,UC-MSCs have this immunomodulation too,but the effects decreased. The platform stage of UC-MSCs was in advance and decurtated after culturing with proper concentration of CoCl2(≤150μmol/L), and the proliferation decreased after 3 days,while the higher concentration of CoCl2 (200、250μmol/L) had a negative impact on the vitality of UC-MSCs.The hypoxic UC-MSCs could inhibit the secretion of IFN-y(compared with controls, P<0.05), while enhanced the secretion of IL-10(compared with controls, P<0.05), however, compared with non-hypoxia group, the secretion of IFN-y increased, IL-10 decreased.
Conclusions The concentration of CoCl2(≤150μmol/L) hardly affected the vitality of UC-MSCs, so it can served as a method of hypoxia.UC-MSCs can inhibited the secretion of IFN-γand promoted the secretion of IL-10 in the immune response of UCB CD34+ progenitor cells, and partly depended on the contacts between cells. Hypoxic UC-MSCs also can inhibit the immune responses of UCB CD34+ progenitor cells, however, the effects decrease compared with normoxia. Co-transplantation of UC-MSCs and UBC CD34+ could can serve as alternative of stem cells transplantation for ischemic cardiomyopathy.
方法通过胶原酶及胰酶消化法从脐带中分离获得MSCs,观察其生长形态,取传代培养至第3代的UC-MSCs流式细胞术检测其表面标志；淋巴细胞分离液分离外周血单个核细胞(PBMC);免疫磁珠分离脐血CD34+祖细胞；将不同数量的第三代UC-MSCs5×103、2×104、8×104分别加入脐血CD34+祖细胞、外周血单个核细胞共培养体系中,不加UC-MSCs的CD34+祖细胞、PBMC共培养体系为阴性对照；另设用Transwel1悬挂式小室将UC-MSCs和脐血CD34+祖细胞、PBMC共培养体系分隔培养；培养96 h后,分别收集上清液,ELISA检测IFN-y和IL-10分泌水平。氯化钴(COCl2)进行化学模拟缺氧,在UC-MSCs中加入氯化钴浓度为0、50、100、150、200、250μmol/L的细胞培养液,分别于12 h、1d、2 d、3d、4d、5d、6 d通过MTT法检测各组的OD值,从而确定各组细胞的增殖情况；将氯化钻浓度为150μmol/L的培养液缺氧处理脐带MSCs48h后,加到脐血CD34+祖细胞、PBMC共培养体系中,培养96 h后,收集上清液,ELISA检测IFN-y和IL-10分泌水平。
结果脐带UC-MSCs高表达CD 105、CD90、CD29、CD49,不表达CD40、CD80、CD86、HLA-DR、CD34、CD45;加UC-MSCs组IFN-γ的分泌量减少(与阴性对照组比,P<0.05),IL-10的分泌量增多(与阴性对照组比,P<0.05),而且具有数量依赖性,即数量越多免疫抑制作用越强；Transwell分隔培养组IFN-γ的分泌量减少(与阴性对照组相比,P<0.05),IL-10的分泌量增多(与阴性对照组相比,P<0.05)；但与接触培养组相比,IFN-y的分泌量增多,IL-10的分泌量减少。经氯化钴模拟缺氧处理的UC-MSCs(150μmol/L)其对数生长期提前,对数期缩短,增殖速度加快,第3天之后减慢,提前达平台期,4-5 d后呈逐渐降低趋势,而经较高浓度(200、250μmol/L)CoCl2处理的UC-MSCs生长曲线趋于低平。缺氧的脐带MSCs能抑制共培养体系中淋巴细胞IFN-y的分泌(与阴性对照组比,P<0.05),同时促进IL-10分泌(与阴性对照组比,P<0.05),但与非缺氧脐带MSCs组相比,IFN-y的分泌量增多,IL-10的分泌量减少。
结论通过酶消化法可从脐带中成功分离获得MSCs;氯化钴(浓度≤150μmol/L)对UC-MSCs增殖影响微弱,可用于其模拟缺氧预处理；缺氧／非缺氧UC-MSCs都能抑制脐血CD34+祖细胞的免疫反应中IFN-γ的分泌,同时促进IL-10分泌,并且这种作用部分依赖于细胞间的相互接触；缺氧处理后的UC-MSCs免疫调节功能下调；UC-MSCs与脐血CD34+祖细胞联合移植可有效减轻同种异体干细胞移植后的免疫排斥反应,提高干细胞的生存率,为治疗缺血性心肌病提供有效的方法。
Objective To investigate wheather the hypoxic/non-hypoxic umbilical cord-derived mesenchymal stem cells can inhibit the immune responses of umbilical cord blood (UCB)CD34+ progenitor cells, affecting the secretion of immunomodulattory cytokines.
Methods UC-MSCs were isolated and cultured, the growth morphology were observed. Surface markers of UC-MSCs were detected by flow cytometry. The CD34 +progenitor cells in umbilical blood were isolated by immunomagnetic beads. Peripheral blood mononuclear cells (PBMCs) were isolated from healthy volunteers using Ficoll-Histopaque density gradients from heparinized samples.The different number of UC-MSCs(5×103、2×104、8×104)were added to the co-culture system of UCB CD34+ progenitor cells and peripheral blood mononuclear cells(PBMC),and cultivation of CD34+ cells and PBMC as negative group, in addition, we mixed this three kinds of cells using Transwell,UBC CD34+ cells and PBMC were cultured in the upper separated from UC-MSCs, after culturing for 96 h, supernatants were collected and detected of the IFN-y and IL-10 level by ELISA. Third passage (P3) UC-MSCs were separately cultivated in DMEM/F12 with cobalt chloride 0、50、100、150、200、250μmol/L at 37℃and 5% CO2 for 12h、1d、2d、3d、4d、5d、6d, detected the proliferation and vitality of every groups by MTT assay. The hypoxic UC-MSCs which were cultivated with CoCl2150μmol/L for 48h were added to the co-culture system of UBC CD34+ cells and PBMC, culturing for 96 h, detected cytokines using above method.
Results UC-MSCs did not express CD40, CD80, CD86, HLA-DR, CD34.It can inhibit IFN-y secretion (compare with controls,P<0.05), promote IL-10 secretion(compared with controls,P<0.05) and had a dose-depended. Using Transwell,UC-MSCs have this immunomodulation too,but the effects decreased. The platform stage of UC-MSCs was in advance and decurtated after culturing with proper concentration of CoCl2(≤150μmol/L), and the proliferation decreased after 3 days,while the higher concentration of CoCl2 (200、250μmol/L) had a negative impact on the vitality of UC-MSCs.The hypoxic UC-MSCs could inhibit the secretion of IFN-y(compared with controls, P<0.05), while enhanced the secretion of IL-10(compared with controls, P<0.05), however, compared with non-hypoxia group, the secretion of IFN-y increased, IL-10 decreased.
Conclusions The concentration of CoCl2(≤150μmol/L) hardly affected the vitality of UC-MSCs, so it can served as a method of hypoxia.UC-MSCs can inhibited the secretion of IFN-γand promoted the secretion of IL-10 in the immune response of UCB CD34+ progenitor cells, and partly depended on the contacts between cells. Hypoxic UC-MSCs also can inhibit the immune responses of UCB CD34+ progenitor cells, however, the effects decrease compared with normoxia. Co-transplantation of UC-MSCs and UBC CD34+ could can serve as alternative of stem cells transplantation for ischemic cardiomyopathy.
引文
[1]Cleland JG, Khand A, Clark A:The heart failure epidemic:exactly how big is it? Eur Heart J,2001,22:623-26.
[2]Quaini F, Urbanek K, Beltrami AP et al:Chimerism of the transplanted heart.N Engl J Med,2002,346:5-15.
[3]Dai W, Hale SL, Kloner RA:Stem cell transplantation for the treatment of myoca-rdial infarction. Transpl Immunol,2005,15:91-97.
[4]Soonpaa MH, Koh GY, Klug MG, Field LJ:Formation of nascent intercalated di-sks between grafted fetal cardiomyocytes and host myocardium.Science,1994, 264:98-101.
[5]Laflanune MA,Mury CE et al.Regeneration the heart[J].Nat Biotechnol 2005,23: 845-856.
[6]Eizawa T, Ikeda U, Murakami Y et al:Decrease in circulating endothelial progen-itor cells in patients with stable coronary artery disease.Heart,2004,90:685-86.
[7]Rao MS, Matton MP. Stem cells and aging:expanding the possibilities[J].Mech Ageing Dev,2001,122(7):713-714.
[8]Nagano M, Yamashita T, Hamada H,et al.Identification of functional endothelial progenitor cells suitable for the treatment of ischemic tissue using human:umbilic-al cord blood.Blood,2007,110(1):151-60.
[9]Fan CL, Li Y, Gao PJ, et al.Differentiation of endothelial progenitor cells from h-uman umbilical cord blood CD 34+ cells in vitro.Acta Pharmacol Sin,2003,24 (3):212-218.
[10]FAN CL, LI Y, GAO PJ, et al.Differentiation of endothelial progenitor cells fro-m human umbilical cord blood CD 34+ cells in vitro[J].Acta Pharmacol Sin, 2003,24(3):212-218.
[11]Thomson JA,Itskovitz EJ,Shapim SS,et al.Embryonic stem cell lines derived from human hlastocysts[J]. Science,1998,282:1145-1147.
[12]Dppolito G, Schiller PC,Ricordi C,et al. Age-related osteogenic potential of mesenchymal stromal stem cells from human vertebral bone marrow[J].J Bone Miner Res,1999,14:1115-1119.
[13]Rao MS,Mattson MP. Stem cells and aging:expanding the possibilities[J].Mech Ageing Dev,2001,122(7):713-734.
[14]Wagner W, Wein F, Seckinger A, et al.Comparative characteristics of mesenchymal stem cells from human bone marrow, adipose tissue, and umbilical cord blood [J].Exp Hematol,2005,33:1402-1416.
[15]Friedman R, Betancur M, Boissel L, et al. Umbilical cord mesenchymal stem cells:adjuvants for human cell transplantation [J].Biol Blood Marrow Transplant, 2007,13:1477-1486.
[16]Fong CY, Richards M, Manasi N, et al. Comparative growth behaviour and characterization of stem cells from human Wharton's jelly [J].Reprod Biomed Online,2007,15:708-718.
[17]Titorencu I, Jinga VV, Constantinescu E, et al.differentiation and characterizeati-on of osteoblasts from human BM mesenchymal cells[J].Cytotherapy,2007, 9:682-696.
[18]Wagner W, Wein F, Seckinger A, et al.Comparative characteristics of mesench-ymal stem cells from human bone marrow, adipose tissue, and umbilical cord blood[J].Exp Hematol,2005,33:1402-1416.
[19]Alhadlaq A, Mao JJ.Mesenchymal stem cells:isolation and therapeutics [J].Stem Cells,2004,13(3):436-448.
[20]Beyer Nardi N, da Silva Meirelles L. Mesenchymal stem cells:isolation, in vitro expansion and characterization [J].Handb Exp Pharmacol,2006,174:249-282.
[21]Schrepfer S,Deuse T, Lange C,et al.Simplified protocol to isolate, purify, and culture expand mesenchymal stem cells [J].Stem Cells,2007,16:105-107.
[22]Wang SH, Hung SC,Peng ST, et al.Mesenchymal stem cells in the Wharton's jelly of the Human Umbilical cord [J].Stem cells,2004,22(8):1330-1337.
[23]Zhao WJ, Chen YJ, Liu W. Study of human bone mar row mesenchymal stem cells separation and its biological characteristic[J].China Journal of Modern Medicine,2006,16(18):2744-2748.
[24]Michowitz Y, Goldstein E, Wexler D, et al.Circulating endothelial progenitor cells and clinical outcome in patients with congestive heart failure[J].Heart, 2007,93(9):1046-50.
[25]Jeong JO, Kim MO, Kim H,et al.Dual angiogenic and neurotrophic effects of bone marrow-derived endothelial progenitor cells on diabetic neuropathy. Circulation,2009,119(5):699-708.
[26]Au P, Daheron LM, Duda DG, et al.Differential in vivo potential of endothelial progenitor cells from human umbilical cord blood and adult peripheral blood to form functional long-lasting vessels[J].Blood,2008,111(3):1302-5.
[27]Azarnoush K, Maurel A, Sebbah L, et al.Enhancement of the functional benefits of skeletal myoblast transplantation by means of administration of hypoxia indu- cible factor-1 alpha[J].J Thorac Cardiovasc Surg,2005,130:173-179.
[28]Tse WT,Pendleton J D, Beyer W M, et al.Suppression of allogeneic T-cell proli-feration by human marrow stromal cells:implication in transplantation[J]. Transplantation,2003,75:389-397.
[29]Tyndall A, Walker UA, Cope A, et al.Immunomodulatory properties of mesenc-hymal stem cells:a review based on an interdisciplinary meeting held at the Kennedy Institute of Rheumatology Division, London, UK,31 October 2005. Arthritis Res Ther 2007,9:301.
[30]Togel F, Hu Z, Weiss K, Isaac J, Lange C,Westenfelder C.Administered mesen-chmal stem cells protect against ischemic acute renal failure through differentiat-ion independent mechanisms[J].Am J Physiol Renal Physiol,2005,289:31-42.
[31]Zappia E, Casazza S,Pedemonte E et al.Mesenchymal stem cells ameliorate experimental autoimmune encephalomyelitis inducing T-cell anergy[J].Blood, 2005,106:1755-61.
[32]Du YY,Zhou SH,Zhou T, et al.Immuno-inflammatory regulation effect of mesenchymal stem cell transplantation in a rat model of myocardial infarction. Cytotherapy[J].2008,10(5):469-478.
[33]Bartholomew A, Sturgoen C,Siatskas M,et al.Mesenchymal stem cellssuppress lymphocyte proliferation in vitro and prolong skin graft survival in vivo[J].Exp Hematol,2002,30(1):42-47.
[34]施小凤,傅晋翔,李军,等.间充质干细胞联合输注促进造血干细胞移植后的造血重建[J].中华器官移植杂志,2005,26(7)：411-415.
[35]Oh W, Kim DS,Yang YS,et al.Immunological properties of umbilical cord blood-derived mesenchymal stromal cells[J].CellImmunol,2008,251(2):116-123.
[36]Wang M, Yang Y, Yang Y et al.The immunomodulatory activity of human umbilical cord blood-derived mesenchymal stem cell in vitro[J].Immunology, 2009,126(2):220-232.
[37]Rainsford E, Reen DJ. Interleukin 10, produced in abundance by human newbor-n T cells may be t he regulator of increased tolerance associated wit h cord blood stem cell transplantation[J]. Br J Haematol,2002,116:702-709.
[38]Tse WT, Pendleton JD, Beyer WM, Egalka MC,Guinan EC.Suppression of allogeneic T-cell proliferation by human marrow stromal cells:implications in transplantation[J].Transplantation,2003,75:389-97.
[39]English K; Barry FP;Field Corbett CP et al.IFN-gama and TNF-alpha differe- ntially regulate immunomodulation by murine mesenchymal stem cells[J]. Immunol Lett,2007,110:91-100.
[40]Beltrami AP, Urbanek K, Kajstura J, et al.Evidence that human cardiacytes myocydivide after myocardial infarction[J].New Engl Med,2001,344(23): 1750-1757.
[41]Dib N, Michler RE, Pagani FD et al.Safety and feasibility of autologous myobl-myoblast transplantation in patients with ischemic cardiomyopathy:four-year follow-up[J].Circulation,2005,112:1748-55.
[42]Orlic D, Kajstura J, Chimenti S, et al.Bone marrow cells regenerate infracted m-yocardium[J].Nature,2001,410:701-5.
[43]Amado LC, Saliaris AP, Schuleri KH et al:Cardiac repair with intramyocardial injection of allogeneic mesenchymal stem cells after myocardial infarction[J]. Proc Natl Acad Sci USA,2005,102:11474-79.
[44]Pompilio G, Cannata A, Peccatori F et al:Autologous peripheral blood stem cell transplantation for myocardial regeneration:a novel strategy for cell collection and surgical injection[J].Ann Thorac Surg,2004,78:1808-12.
[45]Uemura R,Xu M,Ahmad N,et al.Bone marrow stem cells prevent left vent ricular remodeling of ischemic heart t hrough paracrine signaling[J].Circ Res, 2006,98(11):1414-1421.
[46]Hu X, Yu SP, Fraser JL et al.Transplantation of hypoxia-preconditioned mesec-ymal stem cells improves infarcted heart function via enhanced survival of imp-lanted cells and angiogenesis[J].J Thorac Cardiovasc Surg,2008,135:799-808.
[47]Pittenger MF, Martin BJ. Mesenchymal stem cells and their potential as cardiac therapeutics[J].Circ Res 2004,95:9-20.
[48]Thomas E Ichim, Fabio Solano, Roberto Brenes et al.Placental mesenchymal and cord blood stem cell therapy for dilated cardiomyopathy[J].Stem Cell Therapy For Cardiomyopathy,2008,16(6):898-905.
[49]Ardyanto TD, Osaki M, Tokuyasu N, et al.CoC12-induced HIF-alphla expressi-n correlates with proliferation and apoptosis in MKN-1 cells:a possible role for the PI3K/Akt pathway[J].Int Jm Onool,2006,29(3):549-555.
[50]Liu X, Kirschenbaum A, Yao S,et al.Upregulation of vascular endothelial growth factor by cobalt chloride-simulated hypoxia is mediated by persistent induction of cyclo-oxygenase-2 in metastatic human prostate cancer cell line [J]. Clin Exp Metastasis,1999,17(8):687-694.
[51]Bunn HF, Poyton RO.Oxygen sensing and molecular adaptation to hypoxia[J] Physiol Rev,1996,76 (3):839-885.
[1]Murry CE. Haematopoietic stem cells do not transdifferentiate into cardiac myocytes in myocardial infarcts[J].Nature,2004,428 (6983):664-668.
[2]Balsam LB.Haematopoietic stem cells adopt mature haematopoietic fates in ischaemic myocardium[J].Nature,2004,428(6983):668-673.
[3]Kaistura J, Rota M, Whang B,et al.Bone marrow cells differentiate in cardiac celll ineages after infarotion independently of cell fusion[J].Cire Res,2005, 96(1):127-137.
[4]Assmus B.Transplantation of progenitor cells and regeneration enhancement in acute myocardial infarction(TOKARE-AMI)[J].Circulation,2002,106(24):3009 3017
[5]Wolert KC.Intracoronary autologous bone-marrow cell transfer after myocardial infarction:the BOOST randomised controlled clinical trial [J].Lancet,2004,364: 141-148.
[6]Kehat I,Kenyagin-Karsenti D,Sinr M et al.Human embryonic stem cells can differentiate into myocytes with structural and functional property of cardiomyocytes[J].J Clin Invest,2001,108:407-414.
[7]Min J-Y, Yang Y, Converso KL, et al.Transplantation of embryonic stem cells improves cardiac function in postinfarcted rats[J].J Appl Physiol,2002,92:288 -296.
[8]Yang Y, Min J Y,Rana J S,et al.VEGF enhances functional improvement of post infarcted hearts by transplantation of ESC-differentiated cells[J].J Appl Physiol, 2002,93(93):1140-1151.
[9]Min J Y,Yang Y, Sullivan MF.Long-term improvement of cardiac function in rats after infarction by transplantation of embryonic stem cells[J].Thorac cardiovsc Surg,2003,125(2):361-369.
[10]Vittet D, Prandini MH, Berthier R, et al.Embryonic stem cells differentiate in vitro to endothelial cells through successive maturation steps[J].Blood,1996,88: 3424-3431.
[11]Yamashita J, Itoh H, Hirashima M, et al.Flkl-positive cells derived from embryonic stem cells serve as vascular progenitors[J].Nature,2000,408:92-96.
[12]Beltrami AP,Barlucchi L, Torella D, et al. Adult cardiac stem cells are multipotent and support myocardial regeneration[J].Cell,2003,114:763-776.
[13]Oh H, Bradfute SB,Gallardo TD, Nakamura T,et al.Cardiac progenitor cells from adult myocardium:homing, differentiation, and fusion after infarction[J]. Proc Natl Acad Sci U S A,2003,100:12313-12318.
[14]Urbanek K, Torella D, Sheikh F, et al.Myocardial regeneration by activation of multipotentcardiac stem cells in ischemic heart failure[J].Proc Natl Acad Sci U S A,2005,102:8692-8697.
[15]Mouquet F, Pfister O, Jain M, et al.Restoration of cardiac progenitor cells after myocardial infarction by self-proliferation and selective homing of bone marrowderived stem cells[J].Circ Res,2005,97:1090-1092.
[16]Siminiak T, Kalmucki P,Kurpisz M.Autologous skeletal myoblasts for myocardial regeneration[J].J Interv Cardiol,2004,17:357-365.
[17]Murry CE, Wiseman RW, Schwartz SM, Hauschka SD.Skeletal myoblast transplantation for repair of myocardial necrosis. J Clin Invest. 1996;98:2512-2523
[18]Hagege AA, Carrion C,Menasche P, et al.Viability and differentiation of autologous skeletal myoblast grafts in ischaemic cardiomyopathy[J].Lancet, 2003,361:491-492
[19]Menasche P, Hagege AA, Vilquin J-T, et al.Autologous skeletal myoblast transplantation for severe postinfarction left ventricular dysfunction[J].J Am Coll Cardiol,2003,41:1078-1083.
[20]Jackson KA, Majka SM, Wang H, et al.Regeneration of ischemic cardiac muscle and vascular endothelium by adult stem cells[J].J Clin Invest,2001, 107:1395-1402.
[21]Orlic D, Kajstura J, Chimenti S,et al.Bone marrow cells regenerate infarcted myocardium[J].Nature,2001,410:701-705.
[22]Orlic D,Kajstura J,Chimenti S,et al.Mobilized bone marrow cells repair the infarcted heart, improving function and survival[J].Proc Natl Acad Sci U S A., 2001,98:10344-10349.
[23]Murry CE, Soonpaa MH, Reinecke H, et al.Haematopoietic stemcells do not transdifferentiate into cardiac myocytes in myocardial infarcts[J].Nature,2004, 428:664-668.
[0024]Kamihata H, Matsubara H, Nishiue T, et al.Implantation of bone marrow mononuclear cells into ischemic myocardium enhances collateral perfusion and regional function via side supply of angioblasts, angiogenic ligands, and cytokines[J].Circulation,2001,104:1046-1052.
[25]Strauer BE, Brehm M, Zeus T, et al.Repair of infarcted myocardium by autologous intracoronary mononuclear bone marrow cell transplantation in humans[J].Circulation,2002,106:1913-1918.
[26]Au P, Daheron LM, Duda DG, et al.Differential in vivo potential of endothelial progenitor cells from human umbilical cord blood and adult peripheral blood to form functional long-lasting vessels[J].Blood,2008,111(3):1302-5.
[27]Ma N, Stamm C,Kaminski A, et al. Human cord blood cells induce angiogenesis following myocardial infarction in NOD/scid-mice[J].Cardiovasc Res,2005, 66:45-54.
[28]Hirata Y, Sata M, Motomura N.et al.Human umbilical cord blood cells improve cardiac function after myocardial infarction[J].Biochem Biophys Res Commun, 2005,327:609-614.
[29]Pittenger MF, Martin BJ.Mesenchymal stem cells and their potential as cardiac therapeutics[J].Circ Res,2004,95:9-20.
[30]Rao MS,Matton MP.Stem cells and aging:expanding the possibilities[J].Mech Ageing Dev,2001,122(7):713-714.
[31]Wagner W, Wein F,Seckinger A, et al.Comparative characteristics of mesench-ymal stem cells from human bone marrow, adipose tissue, and umbilical cord blo-od[J].Exp Hematol,2005,33:1402-1416.
[321 Tse WT,Pendleton J D, Beyer W M, et al.Suooression of allogeneic T-cell proli-feration by human marrow stromal cells:implication in transplantation [J].Trans-plantation,2003,75:389-397.
[33]Tyndall A, Walker UA, Cope A, et al.Immunomodulatory properties of mesenc-hymal stem cells:a review based on an interdisciplinary meeting held at the Ke-nnedy Institute of Rheumatology Division, London, UK,31 October 2005. Arthritis Res Ther,2007,9:301.
[34]Le Blanc K, Tammik C,Rosendahl K et al.HLA expression and immunologic p-roperities of differentiated and undifferentiared mesenchymal stem cells[J].EXP Hem-etol,2003,31:890-896.
[35]Makino S,Fukuda K, Miyoshi S,et al.Cardiomyocytes can be generated from marrow stromal cells in vitro[J].J Clin Invest,1999,103:697-705.
[36]Shake JG, Gruber PJ, Baumgartner WA, et al.Mesenchymal stem cell implantat-ion in a swine myocardial infarct model:engraftment and functional effects[J]. Ann Thoac Surg,2002,73:1919-1925.
[37]Dai W, Hale SL, Martin BJ, et al.Allogeneic mesenchymal stem cell transplant-ation in postinfarcted rat myocardium:short-and long-term effects[J]. Circulation,2005,112:214-223.
[38]Breitbach M, Bostani T, Roell W, et al. Potential risks of bone marrow cell tran-splantation into infarcted hearts[J].Blood,2007,110:1362-1369.
[39]Noiseux N, Gnecchi M, Lopez-Ilasaca M, et al.Mesenchymal stem cells overex-pressing Akt dramatically repair infarcted myocardium and improvecardiac fun-ction despite infrequent cellular fusion or differentiation[J].Mol Ther,2006,14: 840-850.
[40]Silva GV, Litovsky S,Assad JA, et al.Mesenchymal stem cells differentiate into an endothelial phenotype, enhance vascular density, and improve heart function in a canine chronic ischemia model [J].Circulation,2005,111:150-156.
[41]Tomita S,Li RK, Weisel RD, et al.Autologous transplantation of bone marrow cells improves damaged heart function[J]. Circulation,1999,100:247-256.
[42]Toma C,Pittenger MF, Cahill KS,et al.Human mesenchymal stem cells differ-entiate to a cardiomyocyte phenotype in the adult murine heart[J].Circulation, 2002,105:93-98.
[43]Kinnaird T, Stabile E, Burnett MS,et al.Marrow-derived stromal cells express genes encoding a broad spectrum of arteriogenic cytokines and promote in vitro and in vivo arteriogenesis through paracrine mechanisms[J].Circ Res,2004, 94:678-685.
[44]Massimiliano Gnecchi,Huamei He, Nioseux Noeseux et al.Evidence supporting paracrine hypothesis for Akt-modified mesenchymal stem cell-mediated cardiac prot-ection and functional improvement[J].The FASEB J,2006,20:661-668.
[45]Potier E, Ferreira E, Andriamanalijaona R, et al.Hypoxia affects mesenchymal stromal cell osteogenic differentiation and angiogenic factor expression[J].Bone, 2007,40:1078-1087.
[46]Ceradini DJ, Kulkarni AR, Callaghan MJ, et al.Progenitor cell trafficking is reg-ulated by hypoxic gradients through HIF-1 induction of SDF-1[J].Nat Med, 2004,10:858-864.
[47]Mirotsou M, Zhang Z, Deb A, et al.Secreted frizzled related protein 2 (Sfrp2) is the key Akt-mesenchymal stem cell-released paracrine factor mediating myocar-dial survival and repair[J].Proc Natl Acad Sci U S A,2007,104:1643-1648.
[48]Tyndall A, Walker UA, Cope A, et al.Immunomodulatory properties of mesech- ymal stem cells:a review based on an interdisciplinary meeting held at the Ken-nedy Institute of Rheumatology Division, London, UK,31 October2005. Arthritis Res Ther,2007,9:301-309.
[49]Azarnoush K, Maurel A,Sebbah L,et al.Enhancement of the functional benef-it s of skeletal myoblast t ransplantation by means of coadminist ration of hypox-ia 2 inducible factor 1 alpha[J].J Thorac Cardiovasc Surg,2005,130(1): 173-179.
[50]Jun Guo, Guosheng Lin, Cuiyu Bao et al.Insulin-like growth factor 1 improves the efficacy of mesenchymal stem cells transplantation in a rat model of myocar-dial infarction[J].Journal of Biomedical Science,2008,15:89-97.
[51]C.E. Veltman, O.I.I.Soliman, M.L. Geleijnse, W. B.et al.Four-year follow-up of treatment with intramyocardial skeletal myoblasts injection in patients with ischaemic cardiomyopathy[J].Eur Heart J,2008,171:1-11.
[52]Uemura R,Xu M,Ahmad N,et al.Bone marrow stem cells prevent left vent-ricular remodeling of ischemic heart t hrough paracrine signaling[J]. Circ Res, 2006,98(11):1414-1421.
[53]Hu X, Yu SP,Fraser JL, et al.Transplantation ofhypoxia-preconditioned mesen-cymal stem cells improves.infarcted heart function via enhanced survival of im-planted cells and angiogenesis[J].J Thorac Cardiovasc Surg,2008,135(4):799-808.
[54]Mangi AA,Noiseux N, Kong D, et al.Mesenchymal stem cell s modified with Akt prevent remodeling and restore performance of infarcted hearts[J].Nat Med, 2003,9(9):1195-1201.
[55]Li W, Ma N, Ong LL et al.Bcl-2 engineered MSCs inhibited apoptosis and imp-roved heart function[J].Stem Cells,2007,25(8):2118-27.
[56]Guo Y, He J, Wu J et al. Locally overexpressing hepatocyte growth factor preve-nts post-ischemic heart failure by inhibition of apoptosis viacalcineurin-mediated pathway and angiogenesis[J].Arch Med Res,2008,39(2):179-88.
[57]Sun L, Cui M, Wang Z, et al.Mesenchymal stem cells modified with angiopoiet-in-1 improve remodeling in a rat model of acute myocardial infarction[J].Bioch-em Biophys Res Commun,2007,357(3):779-847.
[58]Yang J,Zhou W, Zheng W, et al.Effects of myocardial transplantation of marro-w mesenchymal stem cells transfected with vascular endothelial growth factor f-or the improvement of heart function and angiogenesis after myocardial infectio-n[J].Cardiology,2007,107(1):17-29.
[59]Wang Y, Johnsen HE, Mortensen S,et al.Changes in circulating mesenchymal stem cells, stem cell homing factor, and vascular growth factors in patients with acute ST elevation myocardial infarction treated with primary percutaneous coronary intervention[J].Heart,2006,92(6):768-74.
[60]Ma J, Ge J, Zhang S,et al.Time course of myocardial stromal cell-drived factor 1 expression and beneficial effects of intravenously administered bone marrow stem cells in rats with experimental muocardial infarction[J].Basic Res Cardiol, 2005,100:217-223.
[61]Nagava N, Kangawa K, Itoh T. Transplantation of mesenchymal stem cells impr-oves cardiac function in a rat model of dilated cardiomyopathy.Circulation, 2005,112(8):1128-35.
[62]Zhang N, Li J, Luo R et al.Bone marrow mesenchymal stem cells induce angiog-enesis and attenuate the remodeling of diabetic cardiomyopathy[J].Arch Med Res,2008,39(2):179-188.
[63]Okada H, Suzuki J, Futamatsu H et al.Attenuation of autoimmune myocarditis in rats by mesenchymal stem cell transplantation through enhanced expression of hepat-ocyte growth factor[J].Int Heart J,2007,48(5):649-61.
[64]Ratajczak MZ, Majka M, Kucia M,et al.Expression of functional CXCR4 by muscles at ellite cells and secretion of SDF-1 by muscle-derived fibroblasts is associated with the presence of both muscle progenitors in bone marrow and hematopoietic stem/progenitor cells in muscles[J].Stem Cells,2003,21(3): 363-371.
[65]Yamaguchi J, Kusano KF, Masuo O, et al.Stromal cell derived factor-1 effects on ex vivo expanded endothelial progenitor cell recruitment for ischemicn eovascularization[J].Circulation,2003,107(9):1322-1328.
[66]Pillarisetti K, Gupta SK. Cloning and relative expression analysis of rat stromal cell derived factor-1(SDF-1):SDF-1 alpham RNA is selectivelyi nduce di n rat model of myocardial infarction farction[J].Inflammation,2001,25(5):293-300.
[67]De Falco E,Porcelli D,Torella AR,et al.SDF-1 involve ment in endothelial phenotype and ischemia-induced recruitment of bone marrow progenitor cells[J]. Blood,2004,104(12):3472-3482.
[68]Neuhaus T, Stier S,Totzke G, et al.Stromal cell-derived factor 1 alpha (SDF-1 alpha) induces gene-expression of early growth response-1(Egr-1)and VEGF in human arterial Endothelial cells and enhances VEGF induced cell proliferation[J].Cell Prolif,2003,36(2):75-86.
[69]Askari AT, Unzek S,Popovic ZB,et al.Effect of stromal-cell-derived factor 1 on stem cell homing and.tissue regeneration in ischaemic cardiomyopathy[J]. Lancet,2003,362(9385):697-703.
[70]Hu X, Dai S,Wu WJ, et al.Stromal cell derived factor-1 alpha confers protection against myocardial ischemia/reperfusion injury:role of the cardiac stromal cell derived factor-1 alpha CXCR4 axis[J].Circulation,2007,116(6): 654-63.
[71]Yoon YS,Park Js, Tkebuchava T, et al.Unexpected severe calification after tra-nsplantation of bone marrow cells in acute myocardial infarction[J].Circulation, 2004,109(25):3154-3157.
[2]Quaini F, Urbanek K, Beltrami AP et al:Chimerism of the transplanted heart.N Engl J Med,2002,346:5-15.
[3]Dai W, Hale SL, Kloner RA:Stem cell transplantation for the treatment of myoca-rdial infarction. Transpl Immunol,2005,15:91-97.
[4]Soonpaa MH, Koh GY, Klug MG, Field LJ:Formation of nascent intercalated di-sks between grafted fetal cardiomyocytes and host myocardium.Science,1994, 264:98-101.
[5]Laflanune MA,Mury CE et al.Regeneration the heart[J].Nat Biotechnol 2005,23: 845-856.
[6]Eizawa T, Ikeda U, Murakami Y et al:Decrease in circulating endothelial progen-itor cells in patients with stable coronary artery disease.Heart,2004,90:685-86.
[7]Rao MS, Matton MP. Stem cells and aging:expanding the possibilities[J].Mech Ageing Dev,2001,122(7):713-714.
[8]Nagano M, Yamashita T, Hamada H,et al.Identification of functional endothelial progenitor cells suitable for the treatment of ischemic tissue using human:umbilic-al cord blood.Blood,2007,110(1):151-60.
[9]Fan CL, Li Y, Gao PJ, et al.Differentiation of endothelial progenitor cells from h-uman umbilical cord blood CD 34+ cells in vitro.Acta Pharmacol Sin,2003,24 (3):212-218.
[10]FAN CL, LI Y, GAO PJ, et al.Differentiation of endothelial progenitor cells fro-m human umbilical cord blood CD 34+ cells in vitro[J].Acta Pharmacol Sin, 2003,24(3):212-218.
[11]Thomson JA,Itskovitz EJ,Shapim SS,et al.Embryonic stem cell lines derived from human hlastocysts[J]. Science,1998,282:1145-1147.
[12]Dppolito G, Schiller PC,Ricordi C,et al. Age-related osteogenic potential of mesenchymal stromal stem cells from human vertebral bone marrow[J].J Bone Miner Res,1999,14:1115-1119.
[13]Rao MS,Mattson MP. Stem cells and aging:expanding the possibilities[J].Mech Ageing Dev,2001,122(7):713-734.
[14]Wagner W, Wein F, Seckinger A, et al.Comparative characteristics of mesenchymal stem cells from human bone marrow, adipose tissue, and umbilical cord blood [J].Exp Hematol,2005,33:1402-1416.
[15]Friedman R, Betancur M, Boissel L, et al. Umbilical cord mesenchymal stem cells:adjuvants for human cell transplantation [J].Biol Blood Marrow Transplant, 2007,13:1477-1486.
[16]Fong CY, Richards M, Manasi N, et al. Comparative growth behaviour and characterization of stem cells from human Wharton's jelly [J].Reprod Biomed Online,2007,15:708-718.
[17]Titorencu I, Jinga VV, Constantinescu E, et al.differentiation and characterizeati-on of osteoblasts from human BM mesenchymal cells[J].Cytotherapy,2007, 9:682-696.
[18]Wagner W, Wein F, Seckinger A, et al.Comparative characteristics of mesench-ymal stem cells from human bone marrow, adipose tissue, and umbilical cord blood[J].Exp Hematol,2005,33:1402-1416.
[19]Alhadlaq A, Mao JJ.Mesenchymal stem cells:isolation and therapeutics [J].Stem Cells,2004,13(3):436-448.
[20]Beyer Nardi N, da Silva Meirelles L. Mesenchymal stem cells:isolation, in vitro expansion and characterization [J].Handb Exp Pharmacol,2006,174:249-282.
[21]Schrepfer S,Deuse T, Lange C,et al.Simplified protocol to isolate, purify, and culture expand mesenchymal stem cells [J].Stem Cells,2007,16:105-107.
[22]Wang SH, Hung SC,Peng ST, et al.Mesenchymal stem cells in the Wharton's jelly of the Human Umbilical cord [J].Stem cells,2004,22(8):1330-1337.
[23]Zhao WJ, Chen YJ, Liu W. Study of human bone mar row mesenchymal stem cells separation and its biological characteristic[J].China Journal of Modern Medicine,2006,16(18):2744-2748.
[24]Michowitz Y, Goldstein E, Wexler D, et al.Circulating endothelial progenitor cells and clinical outcome in patients with congestive heart failure[J].Heart, 2007,93(9):1046-50.
[25]Jeong JO, Kim MO, Kim H,et al.Dual angiogenic and neurotrophic effects of bone marrow-derived endothelial progenitor cells on diabetic neuropathy. Circulation,2009,119(5):699-708.
[26]Au P, Daheron LM, Duda DG, et al.Differential in vivo potential of endothelial progenitor cells from human umbilical cord blood and adult peripheral blood to form functional long-lasting vessels[J].Blood,2008,111(3):1302-5.
[27]Azarnoush K, Maurel A, Sebbah L, et al.Enhancement of the functional benefits of skeletal myoblast transplantation by means of administration of hypoxia indu- cible factor-1 alpha[J].J Thorac Cardiovasc Surg,2005,130:173-179.
[28]Tse WT,Pendleton J D, Beyer W M, et al.Suppression of allogeneic T-cell proli-feration by human marrow stromal cells:implication in transplantation[J]. Transplantation,2003,75:389-397.
[29]Tyndall A, Walker UA, Cope A, et al.Immunomodulatory properties of mesenc-hymal stem cells:a review based on an interdisciplinary meeting held at the Kennedy Institute of Rheumatology Division, London, UK,31 October 2005. Arthritis Res Ther 2007,9:301.
[30]Togel F, Hu Z, Weiss K, Isaac J, Lange C,Westenfelder C.Administered mesen-chmal stem cells protect against ischemic acute renal failure through differentiat-ion independent mechanisms[J].Am J Physiol Renal Physiol,2005,289:31-42.
[31]Zappia E, Casazza S,Pedemonte E et al.Mesenchymal stem cells ameliorate experimental autoimmune encephalomyelitis inducing T-cell anergy[J].Blood, 2005,106:1755-61.
[32]Du YY,Zhou SH,Zhou T, et al.Immuno-inflammatory regulation effect of mesenchymal stem cell transplantation in a rat model of myocardial infarction. Cytotherapy[J].2008,10(5):469-478.
[33]Bartholomew A, Sturgoen C,Siatskas M,et al.Mesenchymal stem cellssuppress lymphocyte proliferation in vitro and prolong skin graft survival in vivo[J].Exp Hematol,2002,30(1):42-47.
[34]施小凤,傅晋翔,李军,等.间充质干细胞联合输注促进造血干细胞移植后的造血重建[J].中华器官移植杂志,2005,26(7)：411-415.
[35]Oh W, Kim DS,Yang YS,et al.Immunological properties of umbilical cord blood-derived mesenchymal stromal cells[J].CellImmunol,2008,251(2):116-123.
[36]Wang M, Yang Y, Yang Y et al.The immunomodulatory activity of human umbilical cord blood-derived mesenchymal stem cell in vitro[J].Immunology, 2009,126(2):220-232.
[37]Rainsford E, Reen DJ. Interleukin 10, produced in abundance by human newbor-n T cells may be t he regulator of increased tolerance associated wit h cord blood stem cell transplantation[J]. Br J Haematol,2002,116:702-709.
[38]Tse WT, Pendleton JD, Beyer WM, Egalka MC,Guinan EC.Suppression of allogeneic T-cell proliferation by human marrow stromal cells:implications in transplantation[J].Transplantation,2003,75:389-97.
[39]English K; Barry FP;Field Corbett CP et al.IFN-gama and TNF-alpha differe- ntially regulate immunomodulation by murine mesenchymal stem cells[J]. Immunol Lett,2007,110:91-100.
[40]Beltrami AP, Urbanek K, Kajstura J, et al.Evidence that human cardiacytes myocydivide after myocardial infarction[J].New Engl Med,2001,344(23): 1750-1757.
[41]Dib N, Michler RE, Pagani FD et al.Safety and feasibility of autologous myobl-myoblast transplantation in patients with ischemic cardiomyopathy:four-year follow-up[J].Circulation,2005,112:1748-55.
[42]Orlic D, Kajstura J, Chimenti S, et al.Bone marrow cells regenerate infracted m-yocardium[J].Nature,2001,410:701-5.
[43]Amado LC, Saliaris AP, Schuleri KH et al:Cardiac repair with intramyocardial injection of allogeneic mesenchymal stem cells after myocardial infarction[J]. Proc Natl Acad Sci USA,2005,102:11474-79.
[44]Pompilio G, Cannata A, Peccatori F et al:Autologous peripheral blood stem cell transplantation for myocardial regeneration:a novel strategy for cell collection and surgical injection[J].Ann Thorac Surg,2004,78:1808-12.
[45]Uemura R,Xu M,Ahmad N,et al.Bone marrow stem cells prevent left vent ricular remodeling of ischemic heart t hrough paracrine signaling[J].Circ Res, 2006,98(11):1414-1421.
[46]Hu X, Yu SP, Fraser JL et al.Transplantation of hypoxia-preconditioned mesec-ymal stem cells improves infarcted heart function via enhanced survival of imp-lanted cells and angiogenesis[J].J Thorac Cardiovasc Surg,2008,135:799-808.
[47]Pittenger MF, Martin BJ. Mesenchymal stem cells and their potential as cardiac therapeutics[J].Circ Res 2004,95:9-20.
[48]Thomas E Ichim, Fabio Solano, Roberto Brenes et al.Placental mesenchymal and cord blood stem cell therapy for dilated cardiomyopathy[J].Stem Cell Therapy For Cardiomyopathy,2008,16(6):898-905.
[49]Ardyanto TD, Osaki M, Tokuyasu N, et al.CoC12-induced HIF-alphla expressi-n correlates with proliferation and apoptosis in MKN-1 cells:a possible role for the PI3K/Akt pathway[J].Int Jm Onool,2006,29(3):549-555.
[50]Liu X, Kirschenbaum A, Yao S,et al.Upregulation of vascular endothelial growth factor by cobalt chloride-simulated hypoxia is mediated by persistent induction of cyclo-oxygenase-2 in metastatic human prostate cancer cell line [J]. Clin Exp Metastasis,1999,17(8):687-694.
[51]Bunn HF, Poyton RO.Oxygen sensing and molecular adaptation to hypoxia[J] Physiol Rev,1996,76 (3):839-885.
[1]Murry CE. Haematopoietic stem cells do not transdifferentiate into cardiac myocytes in myocardial infarcts[J].Nature,2004,428 (6983):664-668.
[2]Balsam LB.Haematopoietic stem cells adopt mature haematopoietic fates in ischaemic myocardium[J].Nature,2004,428(6983):668-673.
[3]Kaistura J, Rota M, Whang B,et al.Bone marrow cells differentiate in cardiac celll ineages after infarotion independently of cell fusion[J].Cire Res,2005, 96(1):127-137.
[4]Assmus B.Transplantation of progenitor cells and regeneration enhancement in acute myocardial infarction(TOKARE-AMI)[J].Circulation,2002,106(24):3009 3017
[5]Wolert KC.Intracoronary autologous bone-marrow cell transfer after myocardial infarction:the BOOST randomised controlled clinical trial [J].Lancet,2004,364: 141-148.
[6]Kehat I,Kenyagin-Karsenti D,Sinr M et al.Human embryonic stem cells can differentiate into myocytes with structural and functional property of cardiomyocytes[J].J Clin Invest,2001,108:407-414.
[7]Min J-Y, Yang Y, Converso KL, et al.Transplantation of embryonic stem cells improves cardiac function in postinfarcted rats[J].J Appl Physiol,2002,92:288 -296.
[8]Yang Y, Min J Y,Rana J S,et al.VEGF enhances functional improvement of post infarcted hearts by transplantation of ESC-differentiated cells[J].J Appl Physiol, 2002,93(93):1140-1151.
[9]Min J Y,Yang Y, Sullivan MF.Long-term improvement of cardiac function in rats after infarction by transplantation of embryonic stem cells[J].Thorac cardiovsc Surg,2003,125(2):361-369.
[10]Vittet D, Prandini MH, Berthier R, et al.Embryonic stem cells differentiate in vitro to endothelial cells through successive maturation steps[J].Blood,1996,88: 3424-3431.
[11]Yamashita J, Itoh H, Hirashima M, et al.Flkl-positive cells derived from embryonic stem cells serve as vascular progenitors[J].Nature,2000,408:92-96.
[12]Beltrami AP,Barlucchi L, Torella D, et al. Adult cardiac stem cells are multipotent and support myocardial regeneration[J].Cell,2003,114:763-776.
[13]Oh H, Bradfute SB,Gallardo TD, Nakamura T,et al.Cardiac progenitor cells from adult myocardium:homing, differentiation, and fusion after infarction[J]. Proc Natl Acad Sci U S A,2003,100:12313-12318.
[14]Urbanek K, Torella D, Sheikh F, et al.Myocardial regeneration by activation of multipotentcardiac stem cells in ischemic heart failure[J].Proc Natl Acad Sci U S A,2005,102:8692-8697.
[15]Mouquet F, Pfister O, Jain M, et al.Restoration of cardiac progenitor cells after myocardial infarction by self-proliferation and selective homing of bone marrowderived stem cells[J].Circ Res,2005,97:1090-1092.
[16]Siminiak T, Kalmucki P,Kurpisz M.Autologous skeletal myoblasts for myocardial regeneration[J].J Interv Cardiol,2004,17:357-365.
[17]Murry CE, Wiseman RW, Schwartz SM, Hauschka SD.Skeletal myoblast transplantation for repair of myocardial necrosis. J Clin Invest. 1996;98:2512-2523
[18]Hagege AA, Carrion C,Menasche P, et al.Viability and differentiation of autologous skeletal myoblast grafts in ischaemic cardiomyopathy[J].Lancet, 2003,361:491-492
[19]Menasche P, Hagege AA, Vilquin J-T, et al.Autologous skeletal myoblast transplantation for severe postinfarction left ventricular dysfunction[J].J Am Coll Cardiol,2003,41:1078-1083.
[20]Jackson KA, Majka SM, Wang H, et al.Regeneration of ischemic cardiac muscle and vascular endothelium by adult stem cells[J].J Clin Invest,2001, 107:1395-1402.
[21]Orlic D, Kajstura J, Chimenti S,et al.Bone marrow cells regenerate infarcted myocardium[J].Nature,2001,410:701-705.
[22]Orlic D,Kajstura J,Chimenti S,et al.Mobilized bone marrow cells repair the infarcted heart, improving function and survival[J].Proc Natl Acad Sci U S A., 2001,98:10344-10349.
[23]Murry CE, Soonpaa MH, Reinecke H, et al.Haematopoietic stemcells do not transdifferentiate into cardiac myocytes in myocardial infarcts[J].Nature,2004, 428:664-668.
[0024]Kamihata H, Matsubara H, Nishiue T, et al.Implantation of bone marrow mononuclear cells into ischemic myocardium enhances collateral perfusion and regional function via side supply of angioblasts, angiogenic ligands, and cytokines[J].Circulation,2001,104:1046-1052.
[25]Strauer BE, Brehm M, Zeus T, et al.Repair of infarcted myocardium by autologous intracoronary mononuclear bone marrow cell transplantation in humans[J].Circulation,2002,106:1913-1918.
[26]Au P, Daheron LM, Duda DG, et al.Differential in vivo potential of endothelial progenitor cells from human umbilical cord blood and adult peripheral blood to form functional long-lasting vessels[J].Blood,2008,111(3):1302-5.
[27]Ma N, Stamm C,Kaminski A, et al. Human cord blood cells induce angiogenesis following myocardial infarction in NOD/scid-mice[J].Cardiovasc Res,2005, 66:45-54.
[28]Hirata Y, Sata M, Motomura N.et al.Human umbilical cord blood cells improve cardiac function after myocardial infarction[J].Biochem Biophys Res Commun, 2005,327:609-614.
[29]Pittenger MF, Martin BJ.Mesenchymal stem cells and their potential as cardiac therapeutics[J].Circ Res,2004,95:9-20.
[30]Rao MS,Matton MP.Stem cells and aging:expanding the possibilities[J].Mech Ageing Dev,2001,122(7):713-714.
[31]Wagner W, Wein F,Seckinger A, et al.Comparative characteristics of mesench-ymal stem cells from human bone marrow, adipose tissue, and umbilical cord blo-od[J].Exp Hematol,2005,33:1402-1416.
[321 Tse WT,Pendleton J D, Beyer W M, et al.Suooression of allogeneic T-cell proli-feration by human marrow stromal cells:implication in transplantation [J].Trans-plantation,2003,75:389-397.
[33]Tyndall A, Walker UA, Cope A, et al.Immunomodulatory properties of mesenc-hymal stem cells:a review based on an interdisciplinary meeting held at the Ke-nnedy Institute of Rheumatology Division, London, UK,31 October 2005. Arthritis Res Ther,2007,9:301.
[34]Le Blanc K, Tammik C,Rosendahl K et al.HLA expression and immunologic p-roperities of differentiated and undifferentiared mesenchymal stem cells[J].EXP Hem-etol,2003,31:890-896.
[35]Makino S,Fukuda K, Miyoshi S,et al.Cardiomyocytes can be generated from marrow stromal cells in vitro[J].J Clin Invest,1999,103:697-705.
[36]Shake JG, Gruber PJ, Baumgartner WA, et al.Mesenchymal stem cell implantat-ion in a swine myocardial infarct model:engraftment and functional effects[J]. Ann Thoac Surg,2002,73:1919-1925.
[37]Dai W, Hale SL, Martin BJ, et al.Allogeneic mesenchymal stem cell transplant-ation in postinfarcted rat myocardium:short-and long-term effects[J]. Circulation,2005,112:214-223.
[38]Breitbach M, Bostani T, Roell W, et al. Potential risks of bone marrow cell tran-splantation into infarcted hearts[J].Blood,2007,110:1362-1369.
[39]Noiseux N, Gnecchi M, Lopez-Ilasaca M, et al.Mesenchymal stem cells overex-pressing Akt dramatically repair infarcted myocardium and improvecardiac fun-ction despite infrequent cellular fusion or differentiation[J].Mol Ther,2006,14: 840-850.
[40]Silva GV, Litovsky S,Assad JA, et al.Mesenchymal stem cells differentiate into an endothelial phenotype, enhance vascular density, and improve heart function in a canine chronic ischemia model [J].Circulation,2005,111:150-156.
[41]Tomita S,Li RK, Weisel RD, et al.Autologous transplantation of bone marrow cells improves damaged heart function[J]. Circulation,1999,100:247-256.
[42]Toma C,Pittenger MF, Cahill KS,et al.Human mesenchymal stem cells differ-entiate to a cardiomyocyte phenotype in the adult murine heart[J].Circulation, 2002,105:93-98.
[43]Kinnaird T, Stabile E, Burnett MS,et al.Marrow-derived stromal cells express genes encoding a broad spectrum of arteriogenic cytokines and promote in vitro and in vivo arteriogenesis through paracrine mechanisms[J].Circ Res,2004, 94:678-685.
[44]Massimiliano Gnecchi,Huamei He, Nioseux Noeseux et al.Evidence supporting paracrine hypothesis for Akt-modified mesenchymal stem cell-mediated cardiac prot-ection and functional improvement[J].The FASEB J,2006,20:661-668.
[45]Potier E, Ferreira E, Andriamanalijaona R, et al.Hypoxia affects mesenchymal stromal cell osteogenic differentiation and angiogenic factor expression[J].Bone, 2007,40:1078-1087.
[46]Ceradini DJ, Kulkarni AR, Callaghan MJ, et al.Progenitor cell trafficking is reg-ulated by hypoxic gradients through HIF-1 induction of SDF-1[J].Nat Med, 2004,10:858-864.
[47]Mirotsou M, Zhang Z, Deb A, et al.Secreted frizzled related protein 2 (Sfrp2) is the key Akt-mesenchymal stem cell-released paracrine factor mediating myocar-dial survival and repair[J].Proc Natl Acad Sci U S A,2007,104:1643-1648.
[48]Tyndall A, Walker UA, Cope A, et al.Immunomodulatory properties of mesech- ymal stem cells:a review based on an interdisciplinary meeting held at the Ken-nedy Institute of Rheumatology Division, London, UK,31 October2005. Arthritis Res Ther,2007,9:301-309.
[49]Azarnoush K, Maurel A,Sebbah L,et al.Enhancement of the functional benef-it s of skeletal myoblast t ransplantation by means of coadminist ration of hypox-ia 2 inducible factor 1 alpha[J].J Thorac Cardiovasc Surg,2005,130(1): 173-179.
[50]Jun Guo, Guosheng Lin, Cuiyu Bao et al.Insulin-like growth factor 1 improves the efficacy of mesenchymal stem cells transplantation in a rat model of myocar-dial infarction[J].Journal of Biomedical Science,2008,15:89-97.
[51]C.E. Veltman, O.I.I.Soliman, M.L. Geleijnse, W. B.et al.Four-year follow-up of treatment with intramyocardial skeletal myoblasts injection in patients with ischaemic cardiomyopathy[J].Eur Heart J,2008,171:1-11.
[52]Uemura R,Xu M,Ahmad N,et al.Bone marrow stem cells prevent left vent-ricular remodeling of ischemic heart t hrough paracrine signaling[J]. Circ Res, 2006,98(11):1414-1421.
[53]Hu X, Yu SP,Fraser JL, et al.Transplantation ofhypoxia-preconditioned mesen-cymal stem cells improves.infarcted heart function via enhanced survival of im-planted cells and angiogenesis[J].J Thorac Cardiovasc Surg,2008,135(4):799-808.
[54]Mangi AA,Noiseux N, Kong D, et al.Mesenchymal stem cell s modified with Akt prevent remodeling and restore performance of infarcted hearts[J].Nat Med, 2003,9(9):1195-1201.
[55]Li W, Ma N, Ong LL et al.Bcl-2 engineered MSCs inhibited apoptosis and imp-roved heart function[J].Stem Cells,2007,25(8):2118-27.
[56]Guo Y, He J, Wu J et al. Locally overexpressing hepatocyte growth factor preve-nts post-ischemic heart failure by inhibition of apoptosis viacalcineurin-mediated pathway and angiogenesis[J].Arch Med Res,2008,39(2):179-88.
[57]Sun L, Cui M, Wang Z, et al.Mesenchymal stem cells modified with angiopoiet-in-1 improve remodeling in a rat model of acute myocardial infarction[J].Bioch-em Biophys Res Commun,2007,357(3):779-847.
[58]Yang J,Zhou W, Zheng W, et al.Effects of myocardial transplantation of marro-w mesenchymal stem cells transfected with vascular endothelial growth factor f-or the improvement of heart function and angiogenesis after myocardial infectio-n[J].Cardiology,2007,107(1):17-29.
[59]Wang Y, Johnsen HE, Mortensen S,et al.Changes in circulating mesenchymal stem cells, stem cell homing factor, and vascular growth factors in patients with acute ST elevation myocardial infarction treated with primary percutaneous coronary intervention[J].Heart,2006,92(6):768-74.
[60]Ma J, Ge J, Zhang S,et al.Time course of myocardial stromal cell-drived factor 1 expression and beneficial effects of intravenously administered bone marrow stem cells in rats with experimental muocardial infarction[J].Basic Res Cardiol, 2005,100:217-223.
[61]Nagava N, Kangawa K, Itoh T. Transplantation of mesenchymal stem cells impr-oves cardiac function in a rat model of dilated cardiomyopathy.Circulation, 2005,112(8):1128-35.
[62]Zhang N, Li J, Luo R et al.Bone marrow mesenchymal stem cells induce angiog-enesis and attenuate the remodeling of diabetic cardiomyopathy[J].Arch Med Res,2008,39(2):179-188.
[63]Okada H, Suzuki J, Futamatsu H et al.Attenuation of autoimmune myocarditis in rats by mesenchymal stem cell transplantation through enhanced expression of hepat-ocyte growth factor[J].Int Heart J,2007,48(5):649-61.
[64]Ratajczak MZ, Majka M, Kucia M,et al.Expression of functional CXCR4 by muscles at ellite cells and secretion of SDF-1 by muscle-derived fibroblasts is associated with the presence of both muscle progenitors in bone marrow and hematopoietic stem/progenitor cells in muscles[J].Stem Cells,2003,21(3): 363-371.
[65]Yamaguchi J, Kusano KF, Masuo O, et al.Stromal cell derived factor-1 effects on ex vivo expanded endothelial progenitor cell recruitment for ischemicn eovascularization[J].Circulation,2003,107(9):1322-1328.
[66]Pillarisetti K, Gupta SK. Cloning and relative expression analysis of rat stromal cell derived factor-1(SDF-1):SDF-1 alpham RNA is selectivelyi nduce di n rat model of myocardial infarction farction[J].Inflammation,2001,25(5):293-300.
[67]De Falco E,Porcelli D,Torella AR,et al.SDF-1 involve ment in endothelial phenotype and ischemia-induced recruitment of bone marrow progenitor cells[J]. Blood,2004,104(12):3472-3482.
[68]Neuhaus T, Stier S,Totzke G, et al.Stromal cell-derived factor 1 alpha (SDF-1 alpha) induces gene-expression of early growth response-1(Egr-1)and VEGF in human arterial Endothelial cells and enhances VEGF induced cell proliferation[J].Cell Prolif,2003,36(2):75-86.
[69]Askari AT, Unzek S,Popovic ZB,et al.Effect of stromal-cell-derived factor 1 on stem cell homing and.tissue regeneration in ischaemic cardiomyopathy[J]. Lancet,2003,362(9385):697-703.
[70]Hu X, Dai S,Wu WJ, et al.Stromal cell derived factor-1 alpha confers protection against myocardial ischemia/reperfusion injury:role of the cardiac stromal cell derived factor-1 alpha CXCR4 axis[J].Circulation,2007,116(6): 654-63.
[71]Yoon YS,Park Js, Tkebuchava T, et al.Unexpected severe calification after tra-nsplantation of bone marrow cells in acute myocardial infarction[J].Circulation, 2004,109(25):3154-3157.