胎鼠脊髓神经干细胞与颌下腺颗粒曲管细胞联合培养的研究

目的研究颌下腺颗粒曲管细胞(GCT细胞)对脊髓神经干细胞(NSCs)增殖分化的影响。方法取胎鼠脊髓NSCs原代和传代培养,取成年雌、雄大鼠GCT细胞原代和传代培养,实验分为3组:NSCs和GCT细胞联合培养组;雌、雄大鼠GCT细胞培养上清液与NSCs培养液混合后培养NSCs;NSCs培养液组为对照组;免疫荧光细胞化学方法进行细胞鉴定,MTT法对比各组NSCs增殖活力。结果NSCs与GCT细胞联合培养7 d后Nestin阳性神经球数多于单独培养,雄性大鼠GCT细胞混合培养液组NSCs增殖活力最强,对照组NSC最弱,雌性混合培养液组介于两者之间。鉴定细胞球Nestin阳性,各组均有少量贴壁分化细胞。结论大鼠GCT细胞分泌物促进NSCs增殖,抑制分化,雄性大鼠GCT细胞作用比雌性大鼠显著。

Co-culture of Granular Convoluted Tubule(GCT) Cells of Rat Submandibular Gland and Neural Stem Cells(NSCs) of Fetal Rat Spinal Cord

Objective To study the effect of GCT cells of rat submandibular gland on the proliferation and differentiation of NSCs of fetal rat spinal cord.Methods NSCs separated from the spinal cord of 13-day old rat fetus were primary and passage cultured in vitro.GCT cells dissociated from adult male and female rats were primary and passage cultured in vitro.NSCs and GCT cells were co-cultured,NSCs were cultured in mingled culture medium(supernatant fluid of GCT cell medium and NSCs culture medium),NSC culture medium was assigned as control group.Immuncytochemical antibody was used to identify the cells,MTT was applied to detect the activity of NSCs.Results More neurospheres expressed Nestin antigen were formed in co-cultured group than in other two groups.NSCs cultured in mingled culture medium of male GCT cells had the highest proliferation ability,NSCs of control group had the minimal proliferation ability,NSCs in mingled culture medium of female GCT cells in between.Neurospheres and small amount of adherent cells were found in each group and all expressed Nestin antigen.Conclusion The proliferation of NSCs was encouraged and the differentiation inhibited by the secretion of GCT cells,the effect is more significant in the GCT cells of male than in those of female.