欢迎加入讨论,如果你接触精原干细胞,即出生后具有self-renew功能的生殖细胞,与前期一直proliferation的PGC不同,它不进行大规模的有丝分裂,而是进行所谓的不对称分裂活动(当然这是在体内in vivo的情况下),在体外培养同样能进行扩增,虽然能传代,但目前并没有建系的报道.关于现成的protocol,我推荐你看humana press的germ cell protocol中的Volume I的Chapter 8:spermatogonial stem cell.上面给出了详细的分离,培养过程.但是我觉得并不十分完美,所以我推荐你看下面的文章 ' K9 V9 h7 s: T1 ]5 M" Q( V6 S4 J2002 Lactate inhibits germ cell apoptosis in the human testis. x6 h5 d' e! f# p6 u7 L
2002 Maintenance of adult mouse type A spermatogonia in vitro influence of serum and growth factors and comparison with prepubertal spermatogonial cell culture : v* v3 }) K7 T& p2 M2003 Clonogenic Culture of Normal Spermatogonia In Vitro Regulation of Postnatal. D3 Y) M- D, c& W- _( M& l$ S0 p+ t1 T
2003 Long-Term Proliferation in Culture and Germline Transmission of Mouse Male Germline Stem Cells4 Q4 t5 @. W7 P: k0 I$ _5 b( [% a
2003 Maintenance of Mouse Male Germ Line Stem Cells In Vitro+ D/ Q5 v8 u, Q
2003 Proliferation and Differentiation of Bovine Type A Spermatogonia During Long Term Culture # U' z% X( K3 K2003 Proliferation and Differentiation of Spermatogonial Stem Cells in the wwv Mutant Mouse Testis' I+ _' r. G* ]/ l& ^% l# e" }
2004 Biological Activity of Cryopreserved Bovine Spermatogonial Stem Cells During In Vitro Culture g' K! \" J2 i: M2 Y
2004 Culture Conditions and Single Growth Factors Affect Fate Determination of Mouse, {/ |1 k a2 Q; p7 j# q
2004 Derivation and morphological characterization of mouse spermatogonial stem cell lines + G+ G9 `3 U0 D2004 Growth factors essential for self-renewal and expansion of mouse spermatogonial stem cells0 J r1 E) U* ]; R
2004 Testis Tissue Explant Culture Supports Survival and Proliferation of Bovine Spermatogonial Stem Cells ( r# {1 o- _& o4 ^( x$ Z% J2005 Anchorage-independent growth of mouse male germline stem cells in vitro R0 N8 f0 f: S2 J
2005 Basic features of bovine spermatogonial culture and effects of glial cell line-derived neurotrophic factor " G5 j- t2 R' z; _& @2005 Conservation of spermatogonial stem cell self-renewal signaling between mouse and rat - _6 M/ d& k8 S# j' X4 |- e; p( f2005 Efforts to create an artificial testis culture systems of male germ cells under biochemical conditions resembling the seminiferous tubular biochemical environment+ B, r9 }3 S3 W$ G" B8 d
2005 Isolation of male germ-line stem cells; influence of GDNF $ F2 O0 P$ c6 @3 w* U2005 Long-Term Culture of Mouse Male Germline Stem Cells Under Serum-or Feeder-Free Conditions " L% E+ p! l. T2005 Self renewal, expansion, and transfection of rat spermatogonial stem cells in culture' K# l8 c( Q: ~# N) [
2006 In vitro propagation of spermatogonial stem cells from KM mice ' A7 Z& Q1 T1 y4 }" R$ ~% [5 M 5 F9 W0 V- w1 {. K! V1 T+ v上面的文章都是成功培养的例子,但是精原干细胞的培养是相当昂贵和耗时的