[关键词]
[摘要]
目的：比较缺氧培养条件下新生大鼠视网膜祖细胞(retinal progenitor cells， RPCs)与脑神经干细胞(neural stem cells， NSCs)钙离子浓度的差异。
方法：分离新生大鼠RPCs及大脑皮质NSCs，进行无血清体外培养； 采用荧光免疫细胞化学的方法进行干细胞鉴定； 缺氧培养后，光镜观察细胞形态，采用钙离子荧光探针结合激光共聚焦显微镜测定细胞内钙离子荧光强度。
结果：缺氧培养后，大脑皮质NSCs较早出现肿胀、伸出突起等变化，随缺氧时间延长，两种细胞内钙离子荧光强度逐渐增强，缺氧12h后RPCs内钙离子荧光强度小于脑皮质NSCs(P<0.05)，差异具有统计学意义。
结论：缺氧培养12h RPCs内钙离子浓度低于大脑皮质NSCs。
[Key word]
[Abstract]
AIM: To compare the difference of the calcium ions concentration between neonatal rats retinal progenitor cells(RPCs)and brain neural stem cells(NSCs)under hypoxia cultivate
in vitro.
METHODS: After isolated from the neonatal SD rats and cultured in serum-free medium in vitro, RPCs and brain NSCs were identified by fluorescent immunocytochemistry. After cultured under hypoxic condition for different time duration, cell morphology were observed under light microscope, and intracellular calcium fluorescence intensity were measured via laser scanning confocal microscopy(LSCM)and calcium fluorescence probe Fluo-4AM.
RESULTS: Cultured under hypoxic conditions, brain NSCs swelled and extended protrusions were noticed earlier than RPCs. With increasing time duration of hypoxia, intracellular calcium ion fluorescence intensity of both types of cells gradually increased. After 12 hours cultured under hypoxic condition, calcium ion fluorescence intensity of RPCs showed significant difference lower than the fluorescence intensity in NSCs(P<0.05).
CONCLUSION: Under hypoxic condition for 12 hours, the concentration of calcium ion in RPCs was lower than that in brain NSCs.
[中图分类号]
[基金项目]
国家自然科学基金资助项目(No. 30772373); 陕西省社发攻关项目(No. 2012K16-11-04); 西安市科技攻关项目(No. GG06173)
