大鼠睾丸支持细胞对培养海马细胞的作用

The Effects of Rat Sertoli Cell on Rat Hippocampal Neurons in Vitro

目的研究睾丸支持细胞对于体外培养大鼠海马细胞的促进生存作用。方法新生SD大鼠海马细胞常规酶解分离培养,12～16dSD大鼠支持细胞采用二步酶消化和低渗分离培养,海马细胞培养分为DMEM组、支持细胞培养液(SCM)组、与支持细胞共培养组。结果SCM组和支持细胞共培养组生存贴壁神经元数量均高于单纯DMEM组,随着培养时间的延长,3组神经元数量均逐渐减少,但神经元的平均突起数量均随时间的延长而增多,且同一时间SCM组和支持细胞共培养组生存神经元数量及平均突起数量均显著高于DMEM组,支持细胞共培养组神经元树突数量较SCM组明显增加。结论支持细胞及其产生的可溶性因子可促进培养的海马神经元的生存和促进突起生长。

Objective To establish the culture systems of sertoli cell（ SC） and hippocampal neurons in vitro,explore the survival and processes growth of neurons with SC co-culture and conditioned culture. Methods Hippocampal neurons of neonatal SD rats were isolated by the enzymatic digestion and cultured in the DMEM media. SC were isolated from 12-16 d SD rats testes by the two-step enzymatic digestion and lower osmosis Tris-HCl treatment. SC culture media were collected and filtered,which was called conditioned media（SCM）. The neurons were cultured with the SC feeder layer and SCM respectively. Results The number of surviving neurons was decreased and the number of processes outgrowth was increased gradually from 1 day to 10 day in the culture. The number of surviving neurons and processes outgrowth in co-culture either with SC group or SCM group were markedly increased than that in control culture group（P〈0.01）.The number of processes outgrowth in co-culture with SC group was markedly increased than that in SCM culture group（P〈0. 01）. Conclusion SC and some soluble factors could secrete can promote hippocampal neurons survival and processes outgrow in vitro.