精原干细胞体外非分化扩增的初步研究

目的探讨精原干细胞体外非分化性扩增的影响因素。方法采用昆明系小白鼠骨髓基质细胞原代培养，待细胞长成单层后经丝裂霉素C处理，作为精原干细胞培养的饲养层，细胞在37℃环境下培养。结果接种后2～4d，精原干细胞数明显增加，细胞单个、成双或成群，非精原干细胞开始出现崩解死亡；接种4～8d，精原干细胞增殖不明显，处于相对静止期；接种10d后，又开始出现明显的分裂增殖，15d已具有精原干细胞特征，继续培养25d，细胞形态未见明显改变，胞质桥仍明显。结论精原干细胞能在37℃环境温度下，在以骨髓基质饲养层上保持良好非分化性扩增。

Proliferation and Undifferentiation of Spermatogonial Stem Cells in vitro

OBJECTIVE: To investigate the fluences on the proliferation and undifferentiation of spermatogonial stem cells in vitro. METHODS: Mouse bone marrow stromal cells were subjected to primary culture; when the cells grew up and extended around, they were treated with mitomycin C. And then, to the bone marrow stromal cells were added spermatogonial stem cells for co-culture. The ambient temperature was 37 degrees C. RESULTS: The spermatogonial stem cells increased in number obviously when cultured for 2-4 days, they could be single, paired or clustered, while the differentiated cells began to die; when cultured for 4-8 days, the cells proliferated not in a distinct manner, they seemed to be stable; but at 10 days, the cells began to grow, especially when cultured for 15 days, the cells had the characteristic structure of the spermatogonial stem cell; when cultured continuously for 25 days, the cells had the same morphology and very clear intercellular bridges. CONCLUSION: Spermatogonial stem cells can proliferate and continue to be undifferentiated on the bone marrow stromal cells when the ambient temperature is 37 degrees C.