摘要
目的比较血清维甲酸(retinoicacid,RA)、胶质细胞源神经营养因子(glialcellline-derivedneurotrophicfactor,GDNF)及脑源性神经营养因子(brainderivedneurotrophicfactor,BDNF)等在不同浓度诱导条件下使恒河猴骨髓基质细胞(bonemarrowstromalcells)诱导分化为神经干细胞(NSCs)及成熟神经细胞的分化条件。方法用Nestin、CD133抗体免疫细胞化学染色,鉴定NSCs;用NSE、-βtublin鉴定神经元;用GFAP鉴定神经胶质细胞,膜片钳检测分化成熟细胞的电生理特性。结果培养第8天多数细胞表现出Nestin及CD133抗原阳性,即为NSCs细胞;诱导后3天即有神经元样细胞出现,此后神经元样细胞逐渐增多,膜片钳检测发现这些细胞具有类似神经细胞的电生理特性。同时,与其他培养条件相比较,低浓度血清(2.5%)+RA+GDNF组诱导分化效能最高。结论应用RA+GDNF及配合使用低浓度血清能够高效诱导骨髓源NSCs向成熟神经细胞分化。
ObjectiveTo explore the conditions to induce differentiation of BMSCs (bone marrow stromal cells)into NSCs(neural stem cells) and mature neural cells. MethodsThe effects of serum, RA, GDNF,BDNF and 2-ME at different concentration were observed. BMSCs were obtained from the marrow of monkey calf and cultured in NSCs culture medium, Nestin, CD133, NSE,β-tublin and GFAP were added as makers to identify the antigen character of cells, and patch-clamp was used to detect the electrophysiological properties of differentiated neural cells. ResultsIt was showed that most cells express the antigen of Nestin and CD133 on 8th day. neuron-like cells can be found after 3 days of induction which have the electrophysiological properties similar to neural cells, RA+GDNF with low concentration of serum was the most effective conditions as compared to other induction conditions in vitro. ConclusionThese results suggested that BMSCs could differentiate into mature neural cells efficiently in vitro in conditions of combining RA+GDNF with low concentration of serum.
出处
《基础医学与临床》
CSCD
北大核心
2005年第5期402-408,共7页
Basic and Clinical Medicine
基金
国家自然科学基金(30270491
30400464)
广东省重大科技项目[粤科基办(2000)25
2004(08)
粤财企(2001)367
(2003)209
粤科计字(2004)112]
军队"十五"攻关课题项目基金(01Z054)
