The Study Of Gene Stra8 And Other Related Genes In The Differentiation Of Msscs In Vitro

Objective:To study the expression of Stra8,mina53,Oct-4 and GCNF in orientation-induced differentiation in mSSCs in vitro, which wouid be useful for learning the optimal condition of culturing mSSCs and the biological characteristics of mSSCs and recognizing the genetic mechanisim of the differentiation in mSSCs.Methods:1,Separated and cultured spermatogonial stem cells of testis of mice aged 6-8 days in vitro. Induced mSSCs differentiate into spermatocyte orientation in vitro by ATRA.2,Markers(c-kit,Oct-4,GCNF) were detected by indirect immunofluorescene in mSSCs before and after induction..3,Spermatogonial stem cells were induced by ATRA for 2h,12h,72h.Meanwhile the levels of Stra8,mina53,Oct-4 and GCNF mRNA expression were detected by RT-PCR method.Results:1,The morphology change of mSSCs indirect immunoinfluorescence showed cultured mSSCs had been differetioned by the induction of ATRA.2,The levels of stra8 mRNA of induced mSSCs for 2h,12h,3d were higher than those of adult mouse testis and uninduced mSSCs,however,the level that mSSCs had been induced for 12 h was the highest of all,and those of 3d and5d were below to which induced for 12 h.The differences between all groups were statistical (p<0.05).3,The expression of mina53 had been increased after mSSCs were induced.The differnces between the 3d-group and the 5d-group weren't statistical(p>0.05). (4) The expression of Oct-4 in undifferentionated mSSCs was the strongest,it had been degenerated since the mSSCs was induced for 2h,it was nagative when mSSCs was induced for 3d and 5d. (5) The expression of GCNF was strengthened since the mSSCs was induced for 12h by ATRA,but it was far lower than the expression in the adult testis of mice, and it was negative in mSSCs induced for 5d.Conclusion:(1)ATRA might have the capability that induces mSSCs differentiate into spermatocyte orientation in vitro.(23)The gene Stra8 maybe initiate the meiosis of mSSCs in its differentiateion by ATRA,but it is yet uncertain that there could be the correlation beween gene Stra8 and gene mina53 in the differentiateion of mSSCs in vitro while induced by ATRA. (3) The expressions of GCNF and Oct-4 are in inverse correlations in the orientation-induced differentiation in mSSCs in vitro by ATRA.