摘要
[目的]探讨精原干细胞（SSCs）冷冻复苏后与Sertoli支持细胞共培养的增殖分化能力。[方法]以7-8 d龄小鼠为材料,Percoll密度梯度离心,并将分离后的SSCs液氮冷冻,复苏后接种至Sertoli饲养层上,37℃下共培养,在倒置相差显微镜下观察细胞的生长特点,并对培养细胞进行C-Kit受体检测。[结果]增殖细胞的生长特点及形态学特征与文献报道精原干细胞的特征相符合。C-Kit受体显示细胞膜呈强阳性。这些均符合精原干细胞的生物学特征。[结论]冷冻复苏后精原干细胞在未加入细胞因子的Sertoli饲养层中能较长时间生长并分裂增殖。
[Objective]To study the potential of generation and differentiation of spermatogonial stem cells（SSCs） which had been freezed and reanimated with Sertoli cells layer.[Methods]Germ cells were isolated from testes of KM mice aged 7-8 days,and were enriched using percoll grads centrifugation;and were freezed by liquid nitrogen,then reanimated and inoculated to sertoli cells layer which treated with mitomycin C were used as feeder cells,the culture medium was DMEM supplemented with 5% FBS,co-culture was done at 37 ℃.Cell growth was observed under inverted phase contrast microscope.Cultured 15-day cells were treated with C-kit receptor test.[Results]The growth characteristics and morphological characteristics of proliferating cells were accorded with those reported in literatures.C-Kit receptor test showed strong positive cell membrane and negative matrix cell,which met the biological characteristics of SSCs.[Conclusion]SSCs can grow,divide and proliferate in feeder layer without cytokine and primary cultured Sertoli cells.
出处
《预防医学论坛》
2008年第12期1060-1062,共3页
Preventive Medicine Tribune
关键词
精原干细胞
冷冻
复苏
SERTOLI细胞
Spermatogonial stem cells
Cryopreservation
Reanimation
Sertoli cells
