制备精原干细胞滋养层细胞条件的优化

目的:优化精原干细胞培养滋养层细胞的制备条件。方法:首先根据文献资料报道和实践经验确定丝裂霉素C作用STO细胞的浓度范围和时间范围,利用双因素优选法缩短丝裂霉素C处理STO细胞的试验范围。然后采用MTT检测法确定丝裂霉素C处理STO细胞的最佳浓度和时间。结果:滋养层细胞处理后不经过冷冻保存的情况下,17.64μg/ml 2 h处理条件的STO细胞在培养14天内细胞数量基本保持稳定,其它处理条件的细胞数量均有所增加;经冷冻保存的情况下,14.72μg/ml 2 h处理条件的STO细胞在培养14天内细胞数量基本保持稳定,其它处理条件的细胞数目均有所降低。结论:滋养层细胞处理后不经过冷冻保存的情况下,17.64μg/ml 2 h的处理条件是丝裂霉素C处理STO细胞的最佳条件;经冷冻保存的情况下,14.72μg/ml 2 h的处理条件是丝裂霉素C处理STO细胞的最佳条件。

Optimization of Preparing Feeder Layer Cells of Spermatogonial Stem Cells

Obejective: To investigate the optimal conditions in establishing STO as spermatogonial stem cells’ feeder layer cells.Methods: According to literature reports and experience,the concentration range and the processing time range of mitomycin C treating STO cells for use as the feeder layers was determined.Two-factor optimization method was used.Two relatively better experimental points that MMC treats STO cells were used.Then,the optimal concentration and treatment time of MMC treating STO cells with MTT method was determined.Results: Without cryopreservation,after treatment with the condition of 17.64μg/ml 2h,the number of STO cells remained stable in culture for 14 days,and the number of STO cells is increased after treatment with the other conditions;With cryopreservation,after treatment with the condition of 14.72 μg/ml 2h,the number of STO cells remained stable in culture for 14 days,and the number of STO cells is decreased after treatment with the other conditions.Conclusion: Without cryopreservation,the condition of 17.64 μg/ml 2h is the optimal condition for mitomycin C treating STO cells;With cryopreservation,the condition of 14.72 μg/ml 2h is the optimal condition for mitomycin C treating STO cells.