制备精原干细胞滋养层细胞条件的优化  

Optimization of Preparing Feeder Layer Cells of Spermatogonial Stem Cells

作　　者：刘海超[1] 张岩[2] 于泊洋[2] 吴应积[1,2] 

机构地区：[1]内蒙古大学生命科学学院,内蒙古呼和浩特010021 [2]内蒙古大学哺乳动物生殖生物学及生物技术教育部重点实验室,内蒙古呼和浩特010021

出　　处：《现代生物医学进展》2012年第4期626-630,共5页Progress in Modern Biomedicine

基　　金：国家基础科学人才培养基金项目(J0730648);教育部"春晖计划"合作科研项目(Z2007-1-01036);高等学校博士学科点专项科研基金项目(20101501110001)

摘　　要：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。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.

关 键 词：精原干细胞 滋养层 丝裂霉素C STO细胞 MTT法 

分 类 号：Q952[生物学—动物学]