摘要
为了研究胶质细胞原性神经营养因子(GDNF)对体外培养小鼠精原干细胞增殖与分化的影响,试验采用差速贴壁法分离小鼠精原干细胞(SSCs)和支持细胞(sertoli),以sertoli为饲养层接种精原干细胞,用免疫荧光染色法对精原干细胞进行鉴定,并将精原干细胞分为2组,试验组培养基中加入GDNF,对照组不加,测定精原干细胞的生长增殖情况,检测精原干细胞生长周期。结果表明:共培养6,9,12,15天时,试验组精原干细胞吸光度值明显高于对照组(P<0.05或0.01)。随着共培养时间的延长,试验组精原干细胞S期染色质含量逐渐增多,然后又逐渐下降,开始另一个分裂周期;与试验组比较,对照组精原干细胞S期染色质含量增长缓慢(P<0.05)。说明GDNF能促进体外分离培养的小鼠精原干细胞的更新增殖。
To study the effect of GNDF on proliferation and differentiation of mouse spermatogonial stem cells in vitro culture. Differential attach- ment method was used to isolate and purify the mouse spermatogonial stem ceils （SSCs） and sertoli cells, taking the sertoli cells as feeder layers for culture of spermatogonial stem cells, and then the spermatogonial stem cells were identified by immunofluorescence staining. The spermatogo- nial stem cells were divided into an experimental group and a control group, and GDNF was only added to the medium of the experimental group. Growth and proliferation of the spermatogonial stem ceils were measured, and the cell cycle of spermatogonial stem cells was determined. The results showed that when the co -cultures were maintained for 6, 9,12, and 15 d, the absorbance value of spermatogonial stem cells in the experimental group was faster than that in the control group（ P 〈 0.05 or 0.01 ）. With prolongation of co - culture time, the content of chroma- tin in spermatogonial stem cells during S phase was increased, and then decreased in the experimental group, then another cell division cycle be- gan. Compared with experimental group , the content of chromatin in spermatogonial stem cells during S phase was slowly increased in the con- trol group（ P 〈 0.05 ）. It indicates that GDNF can promote the proliferation and renewal of spermatogonial stem cells separated in vitro culture.
出处
《黑龙江畜牧兽医》
CAS
北大核心
2012年第10期7-9,161,162,共5页
Heilongjiang Animal Science And veterinary Medicine
基金
山东省自然科学基金项目(ZR2010CL015)
