人羊水干细胞诱导为卵母样生殖细胞的研究
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摘要
将成体干细胞定向诱导分化为卵母样生殖细胞,将会促进卵子发生的相关研究。根据报道,羊水干细胞具有在体外自发分化为包括外胚层、中胚层和内胚层的三个胚层类型细胞的能力。然而,羊水干细胞是否具有发育成生殖细胞的潜能尚未得到证实。在本研究中,我们发现从产前20-38周的人羊水中分离得到一株人羊水干细胞系,在一定的诱导条件下可以分化为卵母样细胞,并且表达多种生殖细胞相关的标记基因。本研究还发现分化过程中形成的少量形态明显特殊的细胞,可进一步发育到更加成熟的生殖样细胞。
实验中采用有限稀释法,从孕中期的羊水中分离纯化了一株羊水干细胞系。染色体核型分析的结果显示其表现为XX型染色体,随后又进一步对该细胞系的特异标志基因进行了鉴定。为了更好地观察诱导细胞在体外的分化及增殖过程,我们在前期工作中构建了pBMP15-EGFP载体。当在添加了猪卵泡液的条件培养基中诱导人羊水干细胞时,我们观察到了一些形态明显特殊的细胞,外面包围着类似透明带样的光环。诱导过程中羊水干细胞可以聚集形成体积较大的悬浮的三维类胚体结构,并且这些类胚体碱性磷酸酶染色呈强阳性。此外在分化过程中,这些细胞还表达很多生殖细胞相关的标志基因,例如Oct4, Dazl, Stella, GDF9, CD117, SCP3 , ZP1, ZP2以及ZP3基因。当转染构建的pBMP15 -EGFP载体时,这些细胞进一步分化成绿色荧光蛋白阳性的卵母样细胞。对羊水干细胞进行进一步诱导后,我们发现其中的一些圆形细胞可以自发分化成类似孤雌囊胚的结构,直径大约在50 - 60μm左右。
将成体干细胞诱导分化为生殖样细胞的研究,尤其是在人羊水干细胞领域进行的相关探索,或许可以提供一个体外的研究模型,来解决早期生殖细胞发育形成中的一些问题。
Derivation of oocyte like cells from adult stem cells would benefit the study of oogenesis. It has been kwon that Human amniotic fluid stem cells (hAFS) are able to differentiate spontaneously in vitro into various somatic cell types. However, the germ cell potential of hAFS cells has never been tested. Here, we report that hAFS cells isolated from the 20 to 38 weeks’gestation have the intrinsic ability to differentiate into oocyte like cells based on several characters specially. The study also tested the hypothesis that a morphologically distinct population of cells derived from hAFS can differentiate further into more mature gametes.
A XX hAFS cell line was isolated from human amniotic fluid of the 2nd trimester of gestation at the single-cell level by limiting dilution, and the specific cell surface antigens of the hAFS cells were characterized. In order to track and identify specific stages of female gamete induction, we construct a pBMP15-EGFP vector. When differentiation was induced in conditioned medium supplemented with porcine follicular fluid, we observed a subpopulation of morphologically round cells with zona pellucida-like morphology were observed which built large embryoid bodies in suspension culture. These 3D embryoid bodies were strongly alkaline phosphatase positive. During differentiation, these cells also express OCT4, Dazl, Stella, GDF9, CD117, SCP3, ZP1, ZP2 and ZP3, which are markers indicative of germ cell formation. When labeled with a pBMP15-EGFP reporter, these cells further differentiate into round GFP-positive oocyte-like cells. Some of these round cells spontaneously developed into parthenogenetic embryo-like structures, which were about 50 - 60μm in diameter.
The potential of generate germ cell precursors from adult stem cells, especially hAFS cells, may provide an in vitro model to study some of the unanswered questions surrounding early germ cell formation.
实验中采用有限稀释法,从孕中期的羊水中分离纯化了一株羊水干细胞系。染色体核型分析的结果显示其表现为XX型染色体,随后又进一步对该细胞系的特异标志基因进行了鉴定。为了更好地观察诱导细胞在体外的分化及增殖过程,我们在前期工作中构建了pBMP15-EGFP载体。当在添加了猪卵泡液的条件培养基中诱导人羊水干细胞时,我们观察到了一些形态明显特殊的细胞,外面包围着类似透明带样的光环。诱导过程中羊水干细胞可以聚集形成体积较大的悬浮的三维类胚体结构,并且这些类胚体碱性磷酸酶染色呈强阳性。此外在分化过程中,这些细胞还表达很多生殖细胞相关的标志基因,例如Oct4, Dazl, Stella, GDF9, CD117, SCP3 , ZP1, ZP2以及ZP3基因。当转染构建的pBMP15 -EGFP载体时,这些细胞进一步分化成绿色荧光蛋白阳性的卵母样细胞。对羊水干细胞进行进一步诱导后,我们发现其中的一些圆形细胞可以自发分化成类似孤雌囊胚的结构,直径大约在50 - 60μm左右。
将成体干细胞诱导分化为生殖样细胞的研究,尤其是在人羊水干细胞领域进行的相关探索,或许可以提供一个体外的研究模型,来解决早期生殖细胞发育形成中的一些问题。
Derivation of oocyte like cells from adult stem cells would benefit the study of oogenesis. It has been kwon that Human amniotic fluid stem cells (hAFS) are able to differentiate spontaneously in vitro into various somatic cell types. However, the germ cell potential of hAFS cells has never been tested. Here, we report that hAFS cells isolated from the 20 to 38 weeks’gestation have the intrinsic ability to differentiate into oocyte like cells based on several characters specially. The study also tested the hypothesis that a morphologically distinct population of cells derived from hAFS can differentiate further into more mature gametes.
A XX hAFS cell line was isolated from human amniotic fluid of the 2nd trimester of gestation at the single-cell level by limiting dilution, and the specific cell surface antigens of the hAFS cells were characterized. In order to track and identify specific stages of female gamete induction, we construct a pBMP15-EGFP vector. When differentiation was induced in conditioned medium supplemented with porcine follicular fluid, we observed a subpopulation of morphologically round cells with zona pellucida-like morphology were observed which built large embryoid bodies in suspension culture. These 3D embryoid bodies were strongly alkaline phosphatase positive. During differentiation, these cells also express OCT4, Dazl, Stella, GDF9, CD117, SCP3, ZP1, ZP2 and ZP3, which are markers indicative of germ cell formation. When labeled with a pBMP15-EGFP reporter, these cells further differentiate into round GFP-positive oocyte-like cells. Some of these round cells spontaneously developed into parthenogenetic embryo-like structures, which were about 50 - 60μm in diameter.
The potential of generate germ cell precursors from adult stem cells, especially hAFS cells, may provide an in vitro model to study some of the unanswered questions surrounding early germ cell formation.
引文
陈陵. 2007.羊水中细胞及其在组织工程领域的研究概况.滨州职业学院学报, 4(4):68-72.
冯建勋,腊晓林,马艳,毕晓娟,温浩. 2009a.两步法分离和培养人羊水来源胚胎间充质干细胞的生物学特征[a].中国组织工程研究与临床康复, 13(45):8854-8860.
冯建勋,腊晓林,马艳,毕晓娟,温浩. 2009b.体外诱导人羊水间充质干细胞向成骨细胞分化.中国误诊学杂志, 10(5): 1012-1017.
冯建勋,腊晓林,马艳,毕晓娟,温浩. 2010. SD大鼠羊水间充质干细胞向神经样细胞体外诱导分化.世界华人消化杂志, 18(3): 250-255.
李宾公,郑泽琪,王梦洪,彭景添,施国祥. 2009.羊水干细胞分泌细胞因子及其对人脐静脉内皮细胞增殖和凋亡的影响.中国组织工程研究与临床康复, 13(45):8849-8853.
李现铎,耿红全,朱哲. 2006.羊水作为间充质干细胞来源的可行性研究.中华小儿外科杂志, 27(9):499-452.
刘慧,李亚里. 2009.羊水来源干细胞的研究进展.现代妇产科进展, 8(3):226-228.
苗竹林. 2003.卵母细胞来源的生长因子GDF-9和GDF-9B/BMP-15.国外医学计划生育分册, 22(4): 204-207.
沈海波,朱哲,潘骏,陈方. 2009.羊水来源胎儿间充质干细胞向成骨细胞的诱导分化.中国组织工程研究与临床康复,13(27):5314-5318.
陶萨茹拉,王峰,刘永斌,呼斯乐,荣威恒. 2009.生长分化因子GDF9基因的研究进展.畜牧与饲料科学, 30(1):54-55.
王晗,陈帅,程祥,窦忠英,王华岩. 2008.通过形成类胚体诱导人羊水多能干细胞向心肌细胞分化.生物工程学报, 24(9): 1582-1587.
王晗,窦忠英,王华岩. 2008.人源羊水干细胞的培养、鉴定及影响因素分析.农业生物技术学报, 16 (5): 804-809.
王启伟,叶玲玲,刘红,吴本传,刘兴茂,李世崇,陈昭烈. 2008.羊水多潜能干细胞的体外培养及其生物学特性[J].中国组织工程研究与临床康复, 12(38):7531-7534.
温叶飞,熊福银,林艳丽,韩正滨,陈红星,邓继先. 2007.小鼠胚胎干细胞向生殖细胞分化的研究.生物技术通讯, 18:700-702.
吴齐全,张胜利,马晓荣,周君梅,陈方. 2009.羊水来源OCT-4阳性胎儿间充质干细胞向血管内皮细胞的体外诱导分化.中国组织工程研究与临床康复, 13(36):7068-7072.
岳小妹,沈雪,李医. 2000.不同胎牛血清培养基对羊水细胞培养结果的影响.山东医药. 49(45):95-96.
郑炜炜,崔光辉,陈系古,韦相才. 2008.小鼠胚胎干细胞体外诱导分化为精原干细胞的研究.生殖与避孕, 28(8): 449-453.
朱哲. 2007.羊水来源胎儿干细胞的研究进展.中华小儿外科杂志, 28(2): 105-107.
Aflatoonian, B, and H Moore. 2006. Germ cells from mouse and human embryonic stem cells. Reproduction, 132:699-707.
Aflatoonian, B, L. Ruban, M Jones, R Aflatoonian, A Fazeli, and HD Moore. 2009. In vitro post-meiotic germ cell development from human embryonic stem cells. Hum Reprod, 24:3150-9.
Barzegari, A, S Atashpaz, K Ghabili, Z Nemati, M Rustaei, and R Azarbaijani. 2009. Polymorphisms in GDF9 and BMP15 Associated with Fertility and Ovulation Rate in Moghani and Ghezel Sheep in Iran.Reprod Domest Anim.
Bossolasco, P, T Montemurro, L Cova, S Zangrossi, C Calzarossa, S Buiatiotis, D Soligo, S Bosari, V Silani, GL Deliliers, P Rebulla, and L Lazzari. 2006. Molecular and phenotypic characterization of human amniotic fluid cells and their differentiation potential. Cell Res, 16:329-36.
Castrillon, D.H., B.J. Quade, T.Y. Wang, C. Quigley, and C.P. Crum. 2000. The human VASA gene is specifically expressed in the germ cell lineage. Proc Natl Acad Sci U S A, 97:9585-90.
Chambers, I, J Silva, D Colby, J Nichols, B Nijmeijer, M Robertson, J Vrana, K Jones, L Grotewold, and A Smith. 2007. Nanog safeguards pluripotency and mediates germline development. Nature, 450:1230-4.
Chen, HF, HC Kuo, CL Chien, CT Shun, YL Yao, PL Ip, CY Chuang, CC. Wang, Y.S. Yang, and H.N. Ho. 2007. Derivation, characterization and differentiation of human embryonic stem cells: comparing serum-containing versus serum-free media and evidence of germ cell differentiation. Hum Reprod, 22:567-77.
Chiavegato, A., S. Bollini, M. Pozzobon, A. Callegari, L. Gasparotto, J. Taiani, M. Piccoli, E. Lenzini, G. Gerosa, I. Vendramin, E. Cozzi, A. Angelini, L. Iop, G.F. Zanon, A. Atala, P. De Coppi, and S. Sartore. 2007. Human amniotic fluid-derived stem cells are rejected after transplantation in the myocardium of normal, ischemic, immuno-suppressed or immuno-deficient rat. J Mol Cell Cardiol, 42:746-59.
Clark, A.T., M.S. Bodnar, M. Fox, R.T. Rodriquez, M.J. Abeyta, M.T. Firpo, and R.A. Pera. 2004. Spontaneous differentiation of germ cells from human embryonic stem cells in vitro. Hum Mol Genet, 13:727-39.
Danner, S., J. Kajahn, C. Geismann, E. Klink, and C. Kruse. 2007. Derivation of oocyte-like cells from a clonal pancreatic stem cell line. Mol Hum Reprod, 13:11-20.
De Coppi, P., G. Bartsch, Jr., M.M. Siddiqui, T. Xu, C.C. Santos, L. Perin, G. Mostoslavsky, A.C. Serre, E.Y. Snyder, J.J. Yoo, M.E. Furth, S. Soker, and A. Atala. 2007. Isolation of amniotic stem cell lines with potential for therapy. Nat Biotechnol, 25:100-6.
DeAngelis, Y.M., C.W. Saunders, K.R. Johnstone, N.L. Reeder, C.G. Coleman, J.R. Kaczvinsky, Jr., C. Gale, R. Walter, M. Mekel, M.P. Lacey, T.W. Keough, A. Fieno, R.A. Grant, B. Begley, Y. Sun, G. Fuentes, R.S. Youngquist, J. Xu, and T.L. Dawson, Jr. 2007. Isolation and expression of a Malassezia globosa lipase gene, LIP1. J Invest Dermatol, 127:2138-46.
Donovan, P.J., and M.P. de Miguel. 2003. Turning germ cells into stem cells. Curr Opin Genet Dev, 13:463-71.
Doyle, D.M., and L.H. Einhorn. 2008. Delayed effects of whole brain radiotherapy in germ cell tumor patients with central nervous system metastases. Int J Radiat Oncol Biol Phys, 70:1361-4.
Dyce, P.W., L. Wen, and J. Li. 2006. In vitro germline potential of stem cells derived from fetal porcine skin. Nat Cell Biol, 8:384-90.
Dyce, P.W., and J. Li. 2006. From skin cells to ovarian follicles? Cell Cycle, 5:1371-5.
Geijsen, N., M. Horoschak, K. Kim, J. Gribnau, K. Eggan, and G.Q. Daley. 2004. Derivation of embryonic germ cells and male gametes from embryonic stem cells. Nature, 427:148-54.
Huang, P., L.M. Lin, X.Y. Wu, Q.L. Tang, X.Y. Feng, G.Y. Lin, X. Lin, H.W. Wang, T.H. Huang, and L. Ma. 2010. Differentiation of human umbilical cord Wharton's jelly-derived mesenchymal stem cells into germ-like cells in vitro. J Cell Biochem, 109:747-54.
Hubner, K., G. Fuhrmann, L.K. Christenson, J. Kehler, R. Reinbold, R. De La Fuente, J. Wood, J.F. Strauss, 3rd, M. Boiani, and H.R. Scholer. 2003. Derivation of oocytes from mouse embryonic stem cells. Science, 300:1251-6.
In't Anker PS, Scherjon SA, Keur CK, Noort WA, Claas FHJ, Willemze R, Fibbe WE, Kanhai H, 2003. Amniotic fluid as a novel source of mesenchymal stem cells for therapeutic transplantation. Blood, 102:1548-1549.
Kaviani A, Perry TE, Dzakovic A, Jennings RW, Ziegler MM, Fauza DO .2001. The amniotic fluid as a source of cells for fetal tissue engineering. J Pediatr Surg, 36: 1662–1665.
Kim, J., Y. Lee, H. Kim, K.J. Hwang, H.C. Kwon, S.K. Kim, D.J. Cho, S.G. Kang, and J. You. 2007. Human amniotic fluid-derived stem cells have characteristics of multipotent stem cells. Cell Prolif, 40:75-90.
Lacham-Kaplan, O., H. Chy, and A. Trounson. 2006. Testicular cell conditioned medium supports differentiation of embryonic stem cells into ovarian structures containing oocytes. Stem Cells, 24:266-73.
Li, C., J. Zhou, G. Shi, Y. Ma, Y. Yang, J. Gu, H. Yu, S. Jin, Z. Wei, F. Chen, and Y. Jin. 2009. Pluripotency can be rapidly and efficiently induced in human amniotic fluid-derived cells. Hum Mol Genet, 18:4340-9.
Linher, K., P. Dyce, and J. Li. 2009. Primordial germ cell-like cells differentiated in vitro from skin-derived stem cells. PLoS One, 4:e8263.
Looijenga LH, Stoop H, de Leeuw HP, de Gouveia Brazao CA, Gillis AJ, van Roozendaal KE, van Zoelen EJ,Weber RF,Wolffenbuttel KP, van Dekken H. 2003. POU5F1 (OCT3/4) identifies cells with pluripotent potential in human germ cell tumors [J]. Cancer Research, 63: 2244–2250.
Lue, Y., K. Erkkila, P.Y. Liu, K. Ma, C. Wang, A.S. Hikim, and R.S. Swerdloff. 2007. Fate of bone marrow stem cells transplanted into the testis: potential implication for men with testicular failure. Am J Pathol, 170:899-908.
Maatouk, D.M., L.D. Kellam, M.R. Mann, H. Lei, E. Li, M.S. Bartolomei, and J.L. Resnick. 2006. DNA methylation is a primary mechanism for silencing postmigratory primordial germ cell genes in both germ cell and somatic cell lineages. Development, 133:3411-8.
Mayanagi, T., K. Ito, and J. Takahashi. 2003. Association of culture of mouse urogenital complexes in media containing rodent sera with the appearance of primordial germ cell-like cells. Reproduction, 125:519-26.
McGuckin, C.P., N. Forraz, M.O. Baradez, S. Navran, J. Zhao, R. Urban, R. Tilton, and L. Denner. 2005. Production of stem cells with embryonic characteristics from human umbilical cord blood. Cell Prolif, 38:245-55.
McLaren, A. 2003. Primordial germ cells in the mouse. Dev Biol, 262:1-15.
Miki, T., T. Lehmann, H. Cai, D.B. Stolz, and S.C. Strom. 2005. Stem cell characteristics of amniotic epithelial cells. Stem Cells, 23:1549-59.
Miki, T., F. Marongiu, E. Ellis, and C.S. S. 2007. Isolation of amniotic epithelial stem cells. Curr Protoc Stem Cell Biol Chapter 1:Unit 1E 3.
Nayernia, K., J.H. Lee, N. Drusenheimer, J. Nolte, G. Wulf, R. Dressel, J. Gromoll, and W. Engel. 2006. Derivation of male germ cells from bone marrow stem cells. Lab Invest, 86:654-63.
Noce, T., S. Okamoto-Ito, and N. Tsunekawa. 2001. Vasa homolog genes in mammalian germ cell development. Cell Struct Funct, 26:131-6.
Old, J.M., N.J. Hill, and E.M. Deane. 2007. Isolation of the mite Myocoptes musculinus Koch from the Spinifex Hopping mouse (Notomys alexis). Lab Anim, 41:292-5.
Peister, A., B.D. Porter, Y.M. Kolambkar, D.W. Hutmacher, and R.E. Guldberg. 2008. Osteogenicdifferentiation of amniotic fluid stem cells. Biomed Mater Eng, 18:241-6.
Pellegrini, M., P. Grimaldi, P. Rossi, R. Geremia, and S. Dolci. 2003. Developmental expression of BMP4/ALK3/SMAD5 signaling pathway in the mouse testis: a potential role of BMP4 in spermatogonia differentiation. J Cell Sci, 116:3363-72.
Pennetier, S., S. Uzbekova, C. Perreau, P. Papillier, P. Mermillod, and R. Dalbies-Tran. 2004. Spatio-temporal expression of the germ cell marker genes MATER, ZAR1, GDF9, BMP15,andVASA in adult bovine tissues, oocytes, and preimplantation embryos. Biol Reprod, 71:1359-66.
Priest RE, Marimuthu KM, Priest JH. 1978, Origin of cells in human amniotic fluid cultures. Lab Invest, 39: 106~109.
Prusa AR, Hengstschl?ger M. 2002. Amniotic fluid cells and human stem cell research: a new connection [J]. Med Sci Monit, 8: 253~257.
Qing, T., Y. Shi, H. Qin, X. Ye, W. Wei, H. Liu, M. Ding, and H. Deng. 2007. Induction of oocyte-like cells from mouse embryonic stem cells by co-culture with ovarian granulosa cells. Differentiation, 75:902-11.
Rajpert-De Meyts E, Bartkova J, Samson M, Hoei-Hansen CE, Frydelund-Larsen L, Bartek J, Skakkebaek NE. 2003. The emerging phenotype of the testicular carcinoma in situ germ cell [J]. Acta Pathologica, Microbiologica, et Immunologica Scandinavica, 111: 267–278.
Richards, A.J., G.C. Enders, and J.L. Resnick. 1999. Differentiation of murine premigratory primordial germ cells in culture. Biol Reprod, 61:1146-51.
Rossi P, Sette C, Dolci S, Geremia R. 2000. Role of c-kit in mammalian spermatogenesis [J]. Journal of Endocrinological Investigation, 23: 609–615.
Salvador, L.M., C.P. Silva, I. Kostetskii, G.L. Radice, and J.F. Strauss, 2008. The promoter of the oocyte-specific gene, Gdf9, is active in population of cultured mouse embryonic stem cells with an oocyte-like phenotype. Methods, 45:172-81.
Schmidt D, Achermann J, Odermatt B, Genoni M, Zund G, Hoerstrup SP. 2008 Jul. Cryopreserved amniotic fluid-derived cells: a lifelong autologous fetal stem cell source for heart valve tissue engineering. J Heart Valve Dis, 17(4):446-55.
Skottman, H., A.M. Stromberg, E. Matilainen, J. Inzunza, O. Hovatta, and R. Lahesmaa. 2006. Unique gene expression signature by human embryonic stem cells cultured under serum-free conditions correlates with their enhanced and prolonged growth in an undifferentiated stage. Stem Cells, 24:151-67.
Stefanidis, K., D. Loutradis, L. Koumbi, V. Anastasiadou, V. Dinopoulou, E. Kiapekou, A.A. Lavdas, S. Mesogitis, and A. Antsaklis. 2008. Deleted in Azoospermia-Like (DAZL) gene-expressing cells in human amniotic fluid: a new source for germ cells research? Fertil Steril, 90:798-804.
Su, Y.Q., X. Wu, M.J. O'Brien, F.L. Pendola, J.N. Denegre, M.M. Matzuk, and J.J. Eppig. 2004. Synergistic roles of BMP15 and GDF9 in the development and function of the oocyte-cumulus cell complex in mice: genetic evidence for an oocyte-granulosa cell regulatory loop. Dev Biol, 276:64-73.
Tilgner, K., S.P. Atkinson, A. Golebiewska, M. Stojkovic, M. Lako, and L. Armstrong. 2008. Isolation of primordial germ cells from differentiating human embryonic stem cells. Stem Cells,26:3075-85.
Toda, A., M. Okabe, T. Yoshida, and T. Nikaido. 2007. The potential of amniotic membrane/amnion-derived cells for regeneration of various tissues. J Pharmacol Sci,105:215-28.
Tsai M S, Hwang S M, Tsai Y L, Cheng F C, Lee J L and Chang Y J. 2006. Clonal amniotic fluid-derived stem cells express characteristics of both mesenchymal and neural stem cells (J). Biology of Reproduction, 74(3): 545~551.
Tsai, M.S., J.L. Lee, Y.J. Chang, and S.M. Hwang. 2004. Isolation of human multipotent mesenchymal stem cells from second-trimester amniotic fluid using a novel two-stage culture protocol. Hum Reprod, 19:1450-6.
Toyooka, Y., N. Tsunekawa, R. Akasu, and T. Noce. 2003. Embryonic stem cells can form germ cells in vitro. Proc Natl Acad Sci U S A, 100:11457-62.
Wei, W., T. Qing, X. Ye, H. Liu, D. Zhang, W. Yang, and H. Deng. 2008. Primordial germ cell specification from embryonic stem cells. PLoS One, 3:e4013.
West, J.A., I.H. Park, G.Q. Daley, and N. Geijsen. 2006. In vitro generation of germ cells from murine embryonic stem cells. Nat Protoc , :2026-36.
West, F.D., D.W. Machacek, N.L. Boyd, K. Pandiyan, K.R. Robbins, and S.L. Stice. 2008. Enrichment and differentiation of human germ-like cells mediated by feeder cells and basic fibroblast growth factor signaling. Stem Cells, 26:2768-76.
Wu, X., and M.M. Matzuk. 2002. GDF-9 and BMP-15: oocyte organizers. Rev Endocr Metab Disord 3:27-32.
Wu, M.Y., and S.N. Chow. 2005. Derivation of germ cells from mouse embryonic stem cells. J Formos Med Assoc, 104:697-706.
Yan, W.H., A. Lin, X.J. Chen, M.Z. Dai, H.H. Xu, B.G. Chen, L.H. Gan, and W.W. Shi. 2008. Immunological aspects of human amniotic fluid cells: implication for normal pregnancy. Cell Biol Int, 32:93-9.
Yu, Z., P. Ji, J. Cao, S. Zhu, Y. Li, L. Zheng, X. Chen, and L. Feng. 2009. Dazl promotes germ cell differentiation from embryonic stem cells. J Mol Cell Biol, 1:93-103.
Zheng, Y.B., Z.L. Gao, C. Xie, H.P. Zhu, L. Peng, J.H. Chen, and Y.T. Chong. 2008. Characterization and hepatogenic differentiation of mesenchymal stem cells from human amniotic fluid and human bone marrow: a comparative study. Cell Biol Int, 32:1439-48.
冯建勋,腊晓林,马艳,毕晓娟,温浩. 2009a.两步法分离和培养人羊水来源胚胎间充质干细胞的生物学特征[a].中国组织工程研究与临床康复, 13(45):8854-8860.
冯建勋,腊晓林,马艳,毕晓娟,温浩. 2009b.体外诱导人羊水间充质干细胞向成骨细胞分化.中国误诊学杂志, 10(5): 1012-1017.
冯建勋,腊晓林,马艳,毕晓娟,温浩. 2010. SD大鼠羊水间充质干细胞向神经样细胞体外诱导分化.世界华人消化杂志, 18(3): 250-255.
李宾公,郑泽琪,王梦洪,彭景添,施国祥. 2009.羊水干细胞分泌细胞因子及其对人脐静脉内皮细胞增殖和凋亡的影响.中国组织工程研究与临床康复, 13(45):8849-8853.
李现铎,耿红全,朱哲. 2006.羊水作为间充质干细胞来源的可行性研究.中华小儿外科杂志, 27(9):499-452.
刘慧,李亚里. 2009.羊水来源干细胞的研究进展.现代妇产科进展, 8(3):226-228.
苗竹林. 2003.卵母细胞来源的生长因子GDF-9和GDF-9B/BMP-15.国外医学计划生育分册, 22(4): 204-207.
沈海波,朱哲,潘骏,陈方. 2009.羊水来源胎儿间充质干细胞向成骨细胞的诱导分化.中国组织工程研究与临床康复,13(27):5314-5318.
陶萨茹拉,王峰,刘永斌,呼斯乐,荣威恒. 2009.生长分化因子GDF9基因的研究进展.畜牧与饲料科学, 30(1):54-55.
王晗,陈帅,程祥,窦忠英,王华岩. 2008.通过形成类胚体诱导人羊水多能干细胞向心肌细胞分化.生物工程学报, 24(9): 1582-1587.
王晗,窦忠英,王华岩. 2008.人源羊水干细胞的培养、鉴定及影响因素分析.农业生物技术学报, 16 (5): 804-809.
王启伟,叶玲玲,刘红,吴本传,刘兴茂,李世崇,陈昭烈. 2008.羊水多潜能干细胞的体外培养及其生物学特性[J].中国组织工程研究与临床康复, 12(38):7531-7534.
温叶飞,熊福银,林艳丽,韩正滨,陈红星,邓继先. 2007.小鼠胚胎干细胞向生殖细胞分化的研究.生物技术通讯, 18:700-702.
吴齐全,张胜利,马晓荣,周君梅,陈方. 2009.羊水来源OCT-4阳性胎儿间充质干细胞向血管内皮细胞的体外诱导分化.中国组织工程研究与临床康复, 13(36):7068-7072.
岳小妹,沈雪,李医. 2000.不同胎牛血清培养基对羊水细胞培养结果的影响.山东医药. 49(45):95-96.
郑炜炜,崔光辉,陈系古,韦相才. 2008.小鼠胚胎干细胞体外诱导分化为精原干细胞的研究.生殖与避孕, 28(8): 449-453.
朱哲. 2007.羊水来源胎儿干细胞的研究进展.中华小儿外科杂志, 28(2): 105-107.
Aflatoonian, B, and H Moore. 2006. Germ cells from mouse and human embryonic stem cells. Reproduction, 132:699-707.
Aflatoonian, B, L. Ruban, M Jones, R Aflatoonian, A Fazeli, and HD Moore. 2009. In vitro post-meiotic germ cell development from human embryonic stem cells. Hum Reprod, 24:3150-9.
Barzegari, A, S Atashpaz, K Ghabili, Z Nemati, M Rustaei, and R Azarbaijani. 2009. Polymorphisms in GDF9 and BMP15 Associated with Fertility and Ovulation Rate in Moghani and Ghezel Sheep in Iran.Reprod Domest Anim.
Bossolasco, P, T Montemurro, L Cova, S Zangrossi, C Calzarossa, S Buiatiotis, D Soligo, S Bosari, V Silani, GL Deliliers, P Rebulla, and L Lazzari. 2006. Molecular and phenotypic characterization of human amniotic fluid cells and their differentiation potential. Cell Res, 16:329-36.
Castrillon, D.H., B.J. Quade, T.Y. Wang, C. Quigley, and C.P. Crum. 2000. The human VASA gene is specifically expressed in the germ cell lineage. Proc Natl Acad Sci U S A, 97:9585-90.
Chambers, I, J Silva, D Colby, J Nichols, B Nijmeijer, M Robertson, J Vrana, K Jones, L Grotewold, and A Smith. 2007. Nanog safeguards pluripotency and mediates germline development. Nature, 450:1230-4.
Chen, HF, HC Kuo, CL Chien, CT Shun, YL Yao, PL Ip, CY Chuang, CC. Wang, Y.S. Yang, and H.N. Ho. 2007. Derivation, characterization and differentiation of human embryonic stem cells: comparing serum-containing versus serum-free media and evidence of germ cell differentiation. Hum Reprod, 22:567-77.
Chiavegato, A., S. Bollini, M. Pozzobon, A. Callegari, L. Gasparotto, J. Taiani, M. Piccoli, E. Lenzini, G. Gerosa, I. Vendramin, E. Cozzi, A. Angelini, L. Iop, G.F. Zanon, A. Atala, P. De Coppi, and S. Sartore. 2007. Human amniotic fluid-derived stem cells are rejected after transplantation in the myocardium of normal, ischemic, immuno-suppressed or immuno-deficient rat. J Mol Cell Cardiol, 42:746-59.
Clark, A.T., M.S. Bodnar, M. Fox, R.T. Rodriquez, M.J. Abeyta, M.T. Firpo, and R.A. Pera. 2004. Spontaneous differentiation of germ cells from human embryonic stem cells in vitro. Hum Mol Genet, 13:727-39.
Danner, S., J. Kajahn, C. Geismann, E. Klink, and C. Kruse. 2007. Derivation of oocyte-like cells from a clonal pancreatic stem cell line. Mol Hum Reprod, 13:11-20.
De Coppi, P., G. Bartsch, Jr., M.M. Siddiqui, T. Xu, C.C. Santos, L. Perin, G. Mostoslavsky, A.C. Serre, E.Y. Snyder, J.J. Yoo, M.E. Furth, S. Soker, and A. Atala. 2007. Isolation of amniotic stem cell lines with potential for therapy. Nat Biotechnol, 25:100-6.
DeAngelis, Y.M., C.W. Saunders, K.R. Johnstone, N.L. Reeder, C.G. Coleman, J.R. Kaczvinsky, Jr., C. Gale, R. Walter, M. Mekel, M.P. Lacey, T.W. Keough, A. Fieno, R.A. Grant, B. Begley, Y. Sun, G. Fuentes, R.S. Youngquist, J. Xu, and T.L. Dawson, Jr. 2007. Isolation and expression of a Malassezia globosa lipase gene, LIP1. J Invest Dermatol, 127:2138-46.
Donovan, P.J., and M.P. de Miguel. 2003. Turning germ cells into stem cells. Curr Opin Genet Dev, 13:463-71.
Doyle, D.M., and L.H. Einhorn. 2008. Delayed effects of whole brain radiotherapy in germ cell tumor patients with central nervous system metastases. Int J Radiat Oncol Biol Phys, 70:1361-4.
Dyce, P.W., L. Wen, and J. Li. 2006. In vitro germline potential of stem cells derived from fetal porcine skin. Nat Cell Biol, 8:384-90.
Dyce, P.W., and J. Li. 2006. From skin cells to ovarian follicles? Cell Cycle, 5:1371-5.
Geijsen, N., M. Horoschak, K. Kim, J. Gribnau, K. Eggan, and G.Q. Daley. 2004. Derivation of embryonic germ cells and male gametes from embryonic stem cells. Nature, 427:148-54.
Huang, P., L.M. Lin, X.Y. Wu, Q.L. Tang, X.Y. Feng, G.Y. Lin, X. Lin, H.W. Wang, T.H. Huang, and L. Ma. 2010. Differentiation of human umbilical cord Wharton's jelly-derived mesenchymal stem cells into germ-like cells in vitro. J Cell Biochem, 109:747-54.
Hubner, K., G. Fuhrmann, L.K. Christenson, J. Kehler, R. Reinbold, R. De La Fuente, J. Wood, J.F. Strauss, 3rd, M. Boiani, and H.R. Scholer. 2003. Derivation of oocytes from mouse embryonic stem cells. Science, 300:1251-6.
In't Anker PS, Scherjon SA, Keur CK, Noort WA, Claas FHJ, Willemze R, Fibbe WE, Kanhai H, 2003. Amniotic fluid as a novel source of mesenchymal stem cells for therapeutic transplantation. Blood, 102:1548-1549.
Kaviani A, Perry TE, Dzakovic A, Jennings RW, Ziegler MM, Fauza DO .2001. The amniotic fluid as a source of cells for fetal tissue engineering. J Pediatr Surg, 36: 1662–1665.
Kim, J., Y. Lee, H. Kim, K.J. Hwang, H.C. Kwon, S.K. Kim, D.J. Cho, S.G. Kang, and J. You. 2007. Human amniotic fluid-derived stem cells have characteristics of multipotent stem cells. Cell Prolif, 40:75-90.
Lacham-Kaplan, O., H. Chy, and A. Trounson. 2006. Testicular cell conditioned medium supports differentiation of embryonic stem cells into ovarian structures containing oocytes. Stem Cells, 24:266-73.
Li, C., J. Zhou, G. Shi, Y. Ma, Y. Yang, J. Gu, H. Yu, S. Jin, Z. Wei, F. Chen, and Y. Jin. 2009. Pluripotency can be rapidly and efficiently induced in human amniotic fluid-derived cells. Hum Mol Genet, 18:4340-9.
Linher, K., P. Dyce, and J. Li. 2009. Primordial germ cell-like cells differentiated in vitro from skin-derived stem cells. PLoS One, 4:e8263.
Looijenga LH, Stoop H, de Leeuw HP, de Gouveia Brazao CA, Gillis AJ, van Roozendaal KE, van Zoelen EJ,Weber RF,Wolffenbuttel KP, van Dekken H. 2003. POU5F1 (OCT3/4) identifies cells with pluripotent potential in human germ cell tumors [J]. Cancer Research, 63: 2244–2250.
Lue, Y., K. Erkkila, P.Y. Liu, K. Ma, C. Wang, A.S. Hikim, and R.S. Swerdloff. 2007. Fate of bone marrow stem cells transplanted into the testis: potential implication for men with testicular failure. Am J Pathol, 170:899-908.
Maatouk, D.M., L.D. Kellam, M.R. Mann, H. Lei, E. Li, M.S. Bartolomei, and J.L. Resnick. 2006. DNA methylation is a primary mechanism for silencing postmigratory primordial germ cell genes in both germ cell and somatic cell lineages. Development, 133:3411-8.
Mayanagi, T., K. Ito, and J. Takahashi. 2003. Association of culture of mouse urogenital complexes in media containing rodent sera with the appearance of primordial germ cell-like cells. Reproduction, 125:519-26.
McGuckin, C.P., N. Forraz, M.O. Baradez, S. Navran, J. Zhao, R. Urban, R. Tilton, and L. Denner. 2005. Production of stem cells with embryonic characteristics from human umbilical cord blood. Cell Prolif, 38:245-55.
McLaren, A. 2003. Primordial germ cells in the mouse. Dev Biol, 262:1-15.
Miki, T., T. Lehmann, H. Cai, D.B. Stolz, and S.C. Strom. 2005. Stem cell characteristics of amniotic epithelial cells. Stem Cells, 23:1549-59.
Miki, T., F. Marongiu, E. Ellis, and C.S. S. 2007. Isolation of amniotic epithelial stem cells. Curr Protoc Stem Cell Biol Chapter 1:Unit 1E 3.
Nayernia, K., J.H. Lee, N. Drusenheimer, J. Nolte, G. Wulf, R. Dressel, J. Gromoll, and W. Engel. 2006. Derivation of male germ cells from bone marrow stem cells. Lab Invest, 86:654-63.
Noce, T., S. Okamoto-Ito, and N. Tsunekawa. 2001. Vasa homolog genes in mammalian germ cell development. Cell Struct Funct, 26:131-6.
Old, J.M., N.J. Hill, and E.M. Deane. 2007. Isolation of the mite Myocoptes musculinus Koch from the Spinifex Hopping mouse (Notomys alexis). Lab Anim, 41:292-5.
Peister, A., B.D. Porter, Y.M. Kolambkar, D.W. Hutmacher, and R.E. Guldberg. 2008. Osteogenicdifferentiation of amniotic fluid stem cells. Biomed Mater Eng, 18:241-6.
Pellegrini, M., P. Grimaldi, P. Rossi, R. Geremia, and S. Dolci. 2003. Developmental expression of BMP4/ALK3/SMAD5 signaling pathway in the mouse testis: a potential role of BMP4 in spermatogonia differentiation. J Cell Sci, 116:3363-72.
Pennetier, S., S. Uzbekova, C. Perreau, P. Papillier, P. Mermillod, and R. Dalbies-Tran. 2004. Spatio-temporal expression of the germ cell marker genes MATER, ZAR1, GDF9, BMP15,andVASA in adult bovine tissues, oocytes, and preimplantation embryos. Biol Reprod, 71:1359-66.
Priest RE, Marimuthu KM, Priest JH. 1978, Origin of cells in human amniotic fluid cultures. Lab Invest, 39: 106~109.
Prusa AR, Hengstschl?ger M. 2002. Amniotic fluid cells and human stem cell research: a new connection [J]. Med Sci Monit, 8: 253~257.
Qing, T., Y. Shi, H. Qin, X. Ye, W. Wei, H. Liu, M. Ding, and H. Deng. 2007. Induction of oocyte-like cells from mouse embryonic stem cells by co-culture with ovarian granulosa cells. Differentiation, 75:902-11.
Rajpert-De Meyts E, Bartkova J, Samson M, Hoei-Hansen CE, Frydelund-Larsen L, Bartek J, Skakkebaek NE. 2003. The emerging phenotype of the testicular carcinoma in situ germ cell [J]. Acta Pathologica, Microbiologica, et Immunologica Scandinavica, 111: 267–278.
Richards, A.J., G.C. Enders, and J.L. Resnick. 1999. Differentiation of murine premigratory primordial germ cells in culture. Biol Reprod, 61:1146-51.
Rossi P, Sette C, Dolci S, Geremia R. 2000. Role of c-kit in mammalian spermatogenesis [J]. Journal of Endocrinological Investigation, 23: 609–615.
Salvador, L.M., C.P. Silva, I. Kostetskii, G.L. Radice, and J.F. Strauss, 2008. The promoter of the oocyte-specific gene, Gdf9, is active in population of cultured mouse embryonic stem cells with an oocyte-like phenotype. Methods, 45:172-81.
Schmidt D, Achermann J, Odermatt B, Genoni M, Zund G, Hoerstrup SP. 2008 Jul. Cryopreserved amniotic fluid-derived cells: a lifelong autologous fetal stem cell source for heart valve tissue engineering. J Heart Valve Dis, 17(4):446-55.
Skottman, H., A.M. Stromberg, E. Matilainen, J. Inzunza, O. Hovatta, and R. Lahesmaa. 2006. Unique gene expression signature by human embryonic stem cells cultured under serum-free conditions correlates with their enhanced and prolonged growth in an undifferentiated stage. Stem Cells, 24:151-67.
Stefanidis, K., D. Loutradis, L. Koumbi, V. Anastasiadou, V. Dinopoulou, E. Kiapekou, A.A. Lavdas, S. Mesogitis, and A. Antsaklis. 2008. Deleted in Azoospermia-Like (DAZL) gene-expressing cells in human amniotic fluid: a new source for germ cells research? Fertil Steril, 90:798-804.
Su, Y.Q., X. Wu, M.J. O'Brien, F.L. Pendola, J.N. Denegre, M.M. Matzuk, and J.J. Eppig. 2004. Synergistic roles of BMP15 and GDF9 in the development and function of the oocyte-cumulus cell complex in mice: genetic evidence for an oocyte-granulosa cell regulatory loop. Dev Biol, 276:64-73.
Tilgner, K., S.P. Atkinson, A. Golebiewska, M. Stojkovic, M. Lako, and L. Armstrong. 2008. Isolation of primordial germ cells from differentiating human embryonic stem cells. Stem Cells,26:3075-85.
Toda, A., M. Okabe, T. Yoshida, and T. Nikaido. 2007. The potential of amniotic membrane/amnion-derived cells for regeneration of various tissues. J Pharmacol Sci,105:215-28.
Tsai M S, Hwang S M, Tsai Y L, Cheng F C, Lee J L and Chang Y J. 2006. Clonal amniotic fluid-derived stem cells express characteristics of both mesenchymal and neural stem cells (J). Biology of Reproduction, 74(3): 545~551.
Tsai, M.S., J.L. Lee, Y.J. Chang, and S.M. Hwang. 2004. Isolation of human multipotent mesenchymal stem cells from second-trimester amniotic fluid using a novel two-stage culture protocol. Hum Reprod, 19:1450-6.
Toyooka, Y., N. Tsunekawa, R. Akasu, and T. Noce. 2003. Embryonic stem cells can form germ cells in vitro. Proc Natl Acad Sci U S A, 100:11457-62.
Wei, W., T. Qing, X. Ye, H. Liu, D. Zhang, W. Yang, and H. Deng. 2008. Primordial germ cell specification from embryonic stem cells. PLoS One, 3:e4013.
West, J.A., I.H. Park, G.Q. Daley, and N. Geijsen. 2006. In vitro generation of germ cells from murine embryonic stem cells. Nat Protoc , :2026-36.
West, F.D., D.W. Machacek, N.L. Boyd, K. Pandiyan, K.R. Robbins, and S.L. Stice. 2008. Enrichment and differentiation of human germ-like cells mediated by feeder cells and basic fibroblast growth factor signaling. Stem Cells, 26:2768-76.
Wu, X., and M.M. Matzuk. 2002. GDF-9 and BMP-15: oocyte organizers. Rev Endocr Metab Disord 3:27-32.
Wu, M.Y., and S.N. Chow. 2005. Derivation of germ cells from mouse embryonic stem cells. J Formos Med Assoc, 104:697-706.
Yan, W.H., A. Lin, X.J. Chen, M.Z. Dai, H.H. Xu, B.G. Chen, L.H. Gan, and W.W. Shi. 2008. Immunological aspects of human amniotic fluid cells: implication for normal pregnancy. Cell Biol Int, 32:93-9.
Yu, Z., P. Ji, J. Cao, S. Zhu, Y. Li, L. Zheng, X. Chen, and L. Feng. 2009. Dazl promotes germ cell differentiation from embryonic stem cells. J Mol Cell Biol, 1:93-103.
Zheng, Y.B., Z.L. Gao, C. Xie, H.P. Zhu, L. Peng, J.H. Chen, and Y.T. Chong. 2008. Characterization and hepatogenic differentiation of mesenchymal stem cells from human amniotic fluid and human bone marrow: a comparative study. Cell Biol Int, 32:1439-48.