精原干细胞体外非分化扩增的初步研究

Proliferation and Undifferentiation of Spermatogonial Stem Cells in vitro

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。

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℃. 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℃.