诱导多能干细胞向雄性生殖细胞分化诱导物的研究进展
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摘要
诱导多能干细胞(induced pluripotent stem cells,iPSCs)是利用细胞重编程技术人工获得的与胚胎干细胞(embryonic stem cells,ESCs)功能类似的细胞,能分化成包括三胚层在内的所有细胞类型,并且规避了ESCs的伦理学争议和移植后的免疫排斥问题,具有十分广阔的应用前景。对iPSCs体外诱导为生殖细胞所用的诱导物及其诱导效果进行了综述,生殖细胞发育机制的研究有望促进未来生殖和发育技术的进步。
Induced pluripotent stem cells (iPSCs) refer to stem cells that are artificially produced by cellular reprogramming,which have similar functions to embryonic stem cells. They can differentiate into all cell types,and avoid the ethical controversy of ESCs and immune rejection after transplantation. They have a broad application prospect. The advances in the in vitro differentiation of male germ cells using i PSCs by different inducers was reviewed,the effect was also investigated. Exploring development mechanisms of germ cells is promising to promote future reproductive and developmental engineering technologies.
Induced pluripotent stem cells (iPSCs) refer to stem cells that are artificially produced by cellular reprogramming,which have similar functions to embryonic stem cells. They can differentiate into all cell types,and avoid the ethical controversy of ESCs and immune rejection after transplantation. They have a broad application prospect. The advances in the in vitro differentiation of male germ cells using i PSCs by different inducers was reviewed,the effect was also investigated. Exploring development mechanisms of germ cells is promising to promote future reproductive and developmental engineering technologies.
引文
[1]Wu Y,Li O,He C,et al.Generation and characterization of induced pluripotent stem cells from guinea pig fetal fibroblasts.Molecular Medicine Reports,2017,15(6):3690-3698.
[2]Wernig M,Zhao J P,Pruszak J,et al.Neurons derived from reprogrammed fibroblasts functionally integrate into the fetal brain and improve symptoms of rats with Parkinson’s disease.Proceedings of National Academy of Sciences of the United States of America,2008,105(15):5856-5861.
[3]Takahashi K,Tanabe K,Ohnuki M,et al.Induction of pluripotent stem cells from human fibroblasts by defined factors.Cell,2007,131(5):861-872.
[4]Imamura M,Aoi T,Tokumasu A,et al.Induction of primordial germ cells from mouse induced pluripotent stem cells derived from adult hepatocytes.Molecular Reproduction and Development,2010,77(9):802-811.
[5]Correia C,Serra M,Espinha N,et al.Combining hypoxia and bioreactor hydrodynamics boosts induced pluripotent stem cell differentiation towards cardiomyocytes.Stem Cell Rev,2014,10(6):786-801.
[6]Schenke-Layland K,Rhodes K E,Angelis E,et al.Reprogrammed mouse fibroblasts differentiate into cells of the cardiovascular and hematopoietic lineages.Stem Cells,2008,26(6):1537-1546.
[7]Shi Y,Inoue H,Wu J C,et al.Induced pluripotent stem cell technology:a decade of progress.Nat Rev Drug Discov,2017,16(2):115-130.
[8]Hikabe O,Hamazaki N,Nagamatsu G,et al.Reconstitution in vitro of the entire cycle of the mouse female germ line.Nature,2016,539(7628):299-303.
[9]Cai H,Xia X,Wang L,et al.In vitro and in vivo differentiation of induced pluripotent stem cells into male germ cells.Biochem Biophys Res Commun,2013,433(3):286-291.
[10]Eguizabal C,Montserrat N,Vassena R,et al.Complete meiosis from human induced pluripotent stem cells.Stem Cells,2011,29(8):1186-1195.
[11]Imamura M,Hikabe O,Lin Z Y C,et al.Generation of germ cells in vitro in the era of induced pluripotent stem cells.Mol Reprod Dev,2014,81(1):2-19.
[12]Yin X H,Li Y,Li J W,et al.Generation and periodontal differentiation of human gingival fibroblasts-derived integrationfree induced pluripotent stem cells.Biochem Biophys Res Commun,2016,473(3):726-732.
[13]Yamanaka S.A fresh look at i PS cells.Cell,2009,137(1):13-17.
[14]Liu S P,Fu R H,Huang Y C,et al.Induced pluripotent stem(i PS)cell research overview.Cell Transplant,2011,20(1):15-19.
[15]Li J,Song W,Pan G,et al.Advances in understanding the cell types and approaches used for generating induced pluripotent stem cells.J Hematol Oncol,2014,7(50):1-18.
[16]Li Y,Wang X,Feng X,et al.Generation of male germ cells from mouse induced pluripotent stem cells in vitro.Stem Cell Res,2014,12(2):517-530
[17]Tan H,Wang J J,Cheng S F,et al.Retinoic acid promotes the proliferation of primordial germ cell-like cells differentiated from mouse skin-derived stem cells in vitro.Theriogenology,2016,85(3):408-418.
[18]Niederreither K,DolléP.Retinoic acid in development:towards an integrated view.Nat Rev Genet,2008,9(7):541-553.
[19]Kim J B,Sebastiano V,Wu G,et al.Oct4-induced pluripotency in adult neural stem cells.Cell,2009,136(3):411-419.
[20]Koubova J,Hu Y C,Bhattacharyya T,et al.Retinoic acid activates two pathways required for meiosis in mice.PLo S Genet,2014,10(8):e1004541.
[21]Costa J J,Souza G B,Soares M A,et al..In vitro differentiation of primordial germ cells and oocyte-like cells from stem cells.Histol Histopathol,2018,33(2):121-132
[22]Morita Y,Tilly J L.Segregation of retinoic acid effects on fetal ovarian germ cell mitosis versus apoptosis by requirement for new macromolecular synthesis.Endocrinology,1999,140(6):2696-2703.
[23]Zhang Y,Wang Y,Zuo Q,et al.Selection of the inducer for the differentiation of chicken embryonic stem cells into male germ cells in vitro.PLo S One,2016,11(10):e0164664.
[24]Geijsen N,Horoschak M,Kim K,et al.Derivation of embryonic germ cells and male gametes from embryonic stem cells.Nature,2004,427(6970):148-154.
[25]Li P,Hu H,Yang S,et al.Differentiation of induced pluripotent stem cells into male germ cells in vitro through embryoid body formation and retinoic acid or testosterone induction.Biomed Res Int,2013,2013(1):608728.
[26]Yang S,Yuan Q,Niu M,et al.BMP4 promotes mouse i PS cell differentiation to male germ cells via Smad1/5,Gata4,Id1 and Id2.Reproduction,2017,153(2):211-220.
[27]Wozney J M.Overview of bone morphogenetic proteins.Spine,2002,27(16):2-8.
[28]He Z.Derivation of male germ cells from induced pluripotent stem(i PS)cells:a novel and crucial source for generating male gametes.Asian J Androl,2012,14(4):516-517.
[29]Lochab A K,Extavour C G.Bone morphogenetic protein(BMP)signaling in animal reproductive system development and function.Dev Biol,2017,427(2):258-269.
[30]Ying Y,Liu X M,Marble A,et al.Requirement of Bmp8b for the generation of primordial germ cells in the mouse.Mol Endocrinol,2000,14(7):1053-1063.
[31]Ying Y,Qi X,Zhao G Q.Induction of primordial germ cells from murine epiblasts by synergistic action of BMP4 and BMP8Bsignaling pathways.Proc Natl Acad Sci USA,2001,98(14):7858-7862.
[32]Kee K,Gonsalves J M,Clark A T,et al.Bone morphogenetic proteins induce germ cell differentiation from human embryonic stem cells.Stem Cells Dev,2006,15(6):831-837.
[33]Panula S,Medrano J V,Kee K,et al.Human germ cell differentiation from fetal-and adult-derived induced pluripotent stem cells.Hum Mol Genet,2011,20(4):752-762.
[34]Bucay N,Yebra M,Cirulli V,et al.A novel approach for the derivation of putative primordial germ cells and sertoli cells from human embryonic stem cells.Stem Cells,2009,27(1):68-77.
[35]Park T S,Galic Z,Conway A E,et al.Derivation of primordial germ cells from human embryonic and induced pluripotent stem cells is significantly improved by coculture with human fetal gonadal cells.Stem Cells,2009,27(4):783-795.
[36]Yu J,Vodyanik M A,Smuga-Otto K,et al.Induced pluripotent stem cell lines derived from human somatic cells.Science,2007,31(5858)8:1917-1920.
[37]Hikabe O,Hamazaki N,Nagamattsu G,et al.Reconstitution in vitro of the entire cycle of the mouse female gern line.Nature.2016,539(7628):299-303.
[38]Hashimoto H,Yuasa S.Testosterone induces cardiomyocyte differentiation from embryonic stem cells.J Mol Cell Cardiol,2013,62(1):69-71.
[39]Silva C,Wood J R,Salvador L,et al.Expression profile of male germ cell-associated genes in mouse embryonic stem cell cultures treated with all-trans retinoic acid and testosterone.Mol Reprod Dev,2009,76(1):11-21.
[40]Narenji Sani R,Tajik P,Movahedin M,et al.Effect of follicle stimulating hormone and testosterone on viability rate of cryopreserved spermatogonial stem cell after thawing.Iran J Vet Sci Technol,2013,5:26-34.
[41]Tajik P,Sani R N,Moezifar M,et al.Effect of folliclestimulating hormone and testosterone on colony formation of bovine spermatogonial stem cell.Comp Clin Pathol 2014,23(4):901-906.
[42]Zanganeh B M,Rastegar T,Roudkenar M H,et al.Co-culture of spermatogonial stem cells with sertoli cells in the presence of testosterone and FSH improved differentiation via up-regulation of post meiotic genes.Acta Med Iran,2013,51(1):1-11.
[43]Silva C,Wood J R,Salvador L,et al.Expression profile of male germ cell-associated genes in mouse embryonic stem cell cultures treated with all-trans retinoic acid and testosterone.Mol Reprod Dev,2009,76(1):11-21.
[44]Correia S,Alves M R,Cavaco J E,et al.Estrogenic regulation of testicular expression of stem cell factor and c-kit:implications in germ cell survival and male fertility.Fertil Steril,2014,102(1):299-306.
[45]Jeong W,Jung S,Bazer F,et al.Stem cell factor-induced AKTcell signaling pathway:effects on porcine trophectoderm and uterine luminal epithelial cells.Gen Comp Endocrinol,2017,250(1):113-121.
[46]Payer B,Saitou M,Barton S C,et al.Stella is a maternal effect gene required for normal early development in mice.Curr Biol,2003,13(23):2110-2117.
[47]Bendall S C,Hughes C,Campbell J L,et al.An enhanced mass spectrometry approach reveals human embryonic stem cell growth factors in culture.Mol Cell Proteomics,2009,8(3):421-432.
[48]West F D,Machacek D W,Boyd N L,et al.Enrichment and differentiation of human germ-like cells mediated by feeder cells and basic fibroblast growth factor signaling.Stem Cells,2008,26(11):2768-2776.
[49]Bahadorani M,Hosseini S M,Abedi P,et al.Glial cell linederived neurotrophic factor in combination with insulin-like growth factor 1 and basic fibroblast growth factor promote in vitro culture of goat spermatogonial stem cells.Growth Factors,2015,33(3):181-191.
[50]Wang H,Xiang J,Zhang W,et al.Induction of germ cell-like cells from porcine induced pluripotent stem cells.Sci Rep,2016,6(1):27256.
[51]Boozarpour S,Matin M M,Momeni-Moghaddam M,et al.Glial cell derived neurotrophic factor induces spermatogonial stem cell marker genes in chicken mesenchymal stem cells.Tissue Cell,2016,48(3):235-241.
[52]Kang L,Wang J,Zhang Y,et al.i PS cells can support full-term development of tetraploid blastocyst-complemented embryos.Cell Stem Cell,2009,5(2):135-138.
[53]Nayernia K,Lee J H,Drusenheimer N,et al.Derivation of male germ cells from bone marrow stem cells.Laboratory Investigation,2006,86(7):654-663.
[54]Dyce P W,Wen L,Li J.In vitro germline potential of stem cells derived from fetal porcine skin.Nature Cell Biology,2006,8(4):384-390.
[55]Zhao X Y,Li W,LüZ,et al.i PS cells produce viable mice through tetraploid complementation.Nature,2009,461(7260):86-90
[56]Kerkis A,Fonseca S A,Serafim R C,et al.In vitro differentiation of male mouse embryonic stem cells into both presumptive sperm cells and oocytes.Cloning Stem Cells,2007,9(4):535-548.
[57]Toyooka Y,Tsunekawa N,Akasu R,et al.Embryonic stem cells can form germ cells in vitro.Proceedings of the National Academy of Sciences,2003,100(20):11457-11462.
[58]Bowles J,Koopman P.Retinoci acid,meiosis and germ cell fate in mammals.Development,2007,134(19):3401-3411.
[59]Bowles J,Knight D,Smith C,et al.Retinoid signaling determines germ cell fate in mice.Science,2006,312(5773):596-600.
[60]Pellegrini M,Grimaldi P,Rossi P,et al.Developmental expression of BMP4/ALK3/SAMD5 signalling pathway in the mouse testis:a potential role of BMP4 in spermatogonia differentiation.Journal of Cell Science,2003,116(16):3363-3372.
[61]West F D,Roche-Rios M I,Abraham S,et al.KIT ligand and bone morphogenetic protein signaling enhances human embryonic stem cell to germ-like cell differentiation.Human Reproduction,2010,25(1):168-178.
[2]Wernig M,Zhao J P,Pruszak J,et al.Neurons derived from reprogrammed fibroblasts functionally integrate into the fetal brain and improve symptoms of rats with Parkinson’s disease.Proceedings of National Academy of Sciences of the United States of America,2008,105(15):5856-5861.
[3]Takahashi K,Tanabe K,Ohnuki M,et al.Induction of pluripotent stem cells from human fibroblasts by defined factors.Cell,2007,131(5):861-872.
[4]Imamura M,Aoi T,Tokumasu A,et al.Induction of primordial germ cells from mouse induced pluripotent stem cells derived from adult hepatocytes.Molecular Reproduction and Development,2010,77(9):802-811.
[5]Correia C,Serra M,Espinha N,et al.Combining hypoxia and bioreactor hydrodynamics boosts induced pluripotent stem cell differentiation towards cardiomyocytes.Stem Cell Rev,2014,10(6):786-801.
[6]Schenke-Layland K,Rhodes K E,Angelis E,et al.Reprogrammed mouse fibroblasts differentiate into cells of the cardiovascular and hematopoietic lineages.Stem Cells,2008,26(6):1537-1546.
[7]Shi Y,Inoue H,Wu J C,et al.Induced pluripotent stem cell technology:a decade of progress.Nat Rev Drug Discov,2017,16(2):115-130.
[8]Hikabe O,Hamazaki N,Nagamatsu G,et al.Reconstitution in vitro of the entire cycle of the mouse female germ line.Nature,2016,539(7628):299-303.
[9]Cai H,Xia X,Wang L,et al.In vitro and in vivo differentiation of induced pluripotent stem cells into male germ cells.Biochem Biophys Res Commun,2013,433(3):286-291.
[10]Eguizabal C,Montserrat N,Vassena R,et al.Complete meiosis from human induced pluripotent stem cells.Stem Cells,2011,29(8):1186-1195.
[11]Imamura M,Hikabe O,Lin Z Y C,et al.Generation of germ cells in vitro in the era of induced pluripotent stem cells.Mol Reprod Dev,2014,81(1):2-19.
[12]Yin X H,Li Y,Li J W,et al.Generation and periodontal differentiation of human gingival fibroblasts-derived integrationfree induced pluripotent stem cells.Biochem Biophys Res Commun,2016,473(3):726-732.
[13]Yamanaka S.A fresh look at i PS cells.Cell,2009,137(1):13-17.
[14]Liu S P,Fu R H,Huang Y C,et al.Induced pluripotent stem(i PS)cell research overview.Cell Transplant,2011,20(1):15-19.
[15]Li J,Song W,Pan G,et al.Advances in understanding the cell types and approaches used for generating induced pluripotent stem cells.J Hematol Oncol,2014,7(50):1-18.
[16]Li Y,Wang X,Feng X,et al.Generation of male germ cells from mouse induced pluripotent stem cells in vitro.Stem Cell Res,2014,12(2):517-530
[17]Tan H,Wang J J,Cheng S F,et al.Retinoic acid promotes the proliferation of primordial germ cell-like cells differentiated from mouse skin-derived stem cells in vitro.Theriogenology,2016,85(3):408-418.
[18]Niederreither K,DolléP.Retinoic acid in development:towards an integrated view.Nat Rev Genet,2008,9(7):541-553.
[19]Kim J B,Sebastiano V,Wu G,et al.Oct4-induced pluripotency in adult neural stem cells.Cell,2009,136(3):411-419.
[20]Koubova J,Hu Y C,Bhattacharyya T,et al.Retinoic acid activates two pathways required for meiosis in mice.PLo S Genet,2014,10(8):e1004541.
[21]Costa J J,Souza G B,Soares M A,et al..In vitro differentiation of primordial germ cells and oocyte-like cells from stem cells.Histol Histopathol,2018,33(2):121-132
[22]Morita Y,Tilly J L.Segregation of retinoic acid effects on fetal ovarian germ cell mitosis versus apoptosis by requirement for new macromolecular synthesis.Endocrinology,1999,140(6):2696-2703.
[23]Zhang Y,Wang Y,Zuo Q,et al.Selection of the inducer for the differentiation of chicken embryonic stem cells into male germ cells in vitro.PLo S One,2016,11(10):e0164664.
[24]Geijsen N,Horoschak M,Kim K,et al.Derivation of embryonic germ cells and male gametes from embryonic stem cells.Nature,2004,427(6970):148-154.
[25]Li P,Hu H,Yang S,et al.Differentiation of induced pluripotent stem cells into male germ cells in vitro through embryoid body formation and retinoic acid or testosterone induction.Biomed Res Int,2013,2013(1):608728.
[26]Yang S,Yuan Q,Niu M,et al.BMP4 promotes mouse i PS cell differentiation to male germ cells via Smad1/5,Gata4,Id1 and Id2.Reproduction,2017,153(2):211-220.
[27]Wozney J M.Overview of bone morphogenetic proteins.Spine,2002,27(16):2-8.
[28]He Z.Derivation of male germ cells from induced pluripotent stem(i PS)cells:a novel and crucial source for generating male gametes.Asian J Androl,2012,14(4):516-517.
[29]Lochab A K,Extavour C G.Bone morphogenetic protein(BMP)signaling in animal reproductive system development and function.Dev Biol,2017,427(2):258-269.
[30]Ying Y,Liu X M,Marble A,et al.Requirement of Bmp8b for the generation of primordial germ cells in the mouse.Mol Endocrinol,2000,14(7):1053-1063.
[31]Ying Y,Qi X,Zhao G Q.Induction of primordial germ cells from murine epiblasts by synergistic action of BMP4 and BMP8Bsignaling pathways.Proc Natl Acad Sci USA,2001,98(14):7858-7862.
[32]Kee K,Gonsalves J M,Clark A T,et al.Bone morphogenetic proteins induce germ cell differentiation from human embryonic stem cells.Stem Cells Dev,2006,15(6):831-837.
[33]Panula S,Medrano J V,Kee K,et al.Human germ cell differentiation from fetal-and adult-derived induced pluripotent stem cells.Hum Mol Genet,2011,20(4):752-762.
[34]Bucay N,Yebra M,Cirulli V,et al.A novel approach for the derivation of putative primordial germ cells and sertoli cells from human embryonic stem cells.Stem Cells,2009,27(1):68-77.
[35]Park T S,Galic Z,Conway A E,et al.Derivation of primordial germ cells from human embryonic and induced pluripotent stem cells is significantly improved by coculture with human fetal gonadal cells.Stem Cells,2009,27(4):783-795.
[36]Yu J,Vodyanik M A,Smuga-Otto K,et al.Induced pluripotent stem cell lines derived from human somatic cells.Science,2007,31(5858)8:1917-1920.
[37]Hikabe O,Hamazaki N,Nagamattsu G,et al.Reconstitution in vitro of the entire cycle of the mouse female gern line.Nature.2016,539(7628):299-303.
[38]Hashimoto H,Yuasa S.Testosterone induces cardiomyocyte differentiation from embryonic stem cells.J Mol Cell Cardiol,2013,62(1):69-71.
[39]Silva C,Wood J R,Salvador L,et al.Expression profile of male germ cell-associated genes in mouse embryonic stem cell cultures treated with all-trans retinoic acid and testosterone.Mol Reprod Dev,2009,76(1):11-21.
[40]Narenji Sani R,Tajik P,Movahedin M,et al.Effect of follicle stimulating hormone and testosterone on viability rate of cryopreserved spermatogonial stem cell after thawing.Iran J Vet Sci Technol,2013,5:26-34.
[41]Tajik P,Sani R N,Moezifar M,et al.Effect of folliclestimulating hormone and testosterone on colony formation of bovine spermatogonial stem cell.Comp Clin Pathol 2014,23(4):901-906.
[42]Zanganeh B M,Rastegar T,Roudkenar M H,et al.Co-culture of spermatogonial stem cells with sertoli cells in the presence of testosterone and FSH improved differentiation via up-regulation of post meiotic genes.Acta Med Iran,2013,51(1):1-11.
[43]Silva C,Wood J R,Salvador L,et al.Expression profile of male germ cell-associated genes in mouse embryonic stem cell cultures treated with all-trans retinoic acid and testosterone.Mol Reprod Dev,2009,76(1):11-21.
[44]Correia S,Alves M R,Cavaco J E,et al.Estrogenic regulation of testicular expression of stem cell factor and c-kit:implications in germ cell survival and male fertility.Fertil Steril,2014,102(1):299-306.
[45]Jeong W,Jung S,Bazer F,et al.Stem cell factor-induced AKTcell signaling pathway:effects on porcine trophectoderm and uterine luminal epithelial cells.Gen Comp Endocrinol,2017,250(1):113-121.
[46]Payer B,Saitou M,Barton S C,et al.Stella is a maternal effect gene required for normal early development in mice.Curr Biol,2003,13(23):2110-2117.
[47]Bendall S C,Hughes C,Campbell J L,et al.An enhanced mass spectrometry approach reveals human embryonic stem cell growth factors in culture.Mol Cell Proteomics,2009,8(3):421-432.
[48]West F D,Machacek D W,Boyd N L,et al.Enrichment and differentiation of human germ-like cells mediated by feeder cells and basic fibroblast growth factor signaling.Stem Cells,2008,26(11):2768-2776.
[49]Bahadorani M,Hosseini S M,Abedi P,et al.Glial cell linederived neurotrophic factor in combination with insulin-like growth factor 1 and basic fibroblast growth factor promote in vitro culture of goat spermatogonial stem cells.Growth Factors,2015,33(3):181-191.
[50]Wang H,Xiang J,Zhang W,et al.Induction of germ cell-like cells from porcine induced pluripotent stem cells.Sci Rep,2016,6(1):27256.
[51]Boozarpour S,Matin M M,Momeni-Moghaddam M,et al.Glial cell derived neurotrophic factor induces spermatogonial stem cell marker genes in chicken mesenchymal stem cells.Tissue Cell,2016,48(3):235-241.
[52]Kang L,Wang J,Zhang Y,et al.i PS cells can support full-term development of tetraploid blastocyst-complemented embryos.Cell Stem Cell,2009,5(2):135-138.
[53]Nayernia K,Lee J H,Drusenheimer N,et al.Derivation of male germ cells from bone marrow stem cells.Laboratory Investigation,2006,86(7):654-663.
[54]Dyce P W,Wen L,Li J.In vitro germline potential of stem cells derived from fetal porcine skin.Nature Cell Biology,2006,8(4):384-390.
[55]Zhao X Y,Li W,LüZ,et al.i PS cells produce viable mice through tetraploid complementation.Nature,2009,461(7260):86-90
[56]Kerkis A,Fonseca S A,Serafim R C,et al.In vitro differentiation of male mouse embryonic stem cells into both presumptive sperm cells and oocytes.Cloning Stem Cells,2007,9(4):535-548.
[57]Toyooka Y,Tsunekawa N,Akasu R,et al.Embryonic stem cells can form germ cells in vitro.Proceedings of the National Academy of Sciences,2003,100(20):11457-11462.
[58]Bowles J,Koopman P.Retinoci acid,meiosis and germ cell fate in mammals.Development,2007,134(19):3401-3411.
[59]Bowles J,Knight D,Smith C,et al.Retinoid signaling determines germ cell fate in mice.Science,2006,312(5773):596-600.
[60]Pellegrini M,Grimaldi P,Rossi P,et al.Developmental expression of BMP4/ALK3/SAMD5 signalling pathway in the mouse testis:a potential role of BMP4 in spermatogonia differentiation.Journal of Cell Science,2003,116(16):3363-3372.
[61]West F D,Roche-Rios M I,Abraham S,et al.KIT ligand and bone morphogenetic protein signaling enhances human embryonic stem cell to germ-like cell differentiation.Human Reproduction,2010,25(1):168-178.