摘要
目的 :从人胚胎脑海马区分离、培养并鉴定神经前体细胞 ,观察其形态学变化。方法 :取胎龄 8～ 12周人胚胎脑海马区细胞 ,用含人表皮生长因子 (h -EGF)、人碱性成纤维细胞生长因子 (h -bFGF)以及人白细胞抑制因子 (h -LIF)的DMEM F12培养液离体培养 ,以 1%胎牛血清 (FBS)和多聚赖氨酸诱导分化 ,倒置显微镜下测量形态指标 ,免疫细胞化学反应分析其神经化学性质。结果 :原代培养中 ,可见许多悬浮生长并渐增大的细胞球 ,以及很少数贴壁生长的成簇或单个细胞。将球吹打传代后 ,又有许多新的细胞球生成。取细胞球诱导分化 ,则见细胞从球迁出、伸出突起并渐分支 ,细胞分别 β -Ⅲtubulin、NF - 2 0 0、GFAP、Galc等免疫细胞化学反应阳性。 结论 :人胚胎脑海马区有神经前体细胞存在 ,离体培养时能分裂增殖和自我更新 ,并能被诱导分化为神经元前体细胞、神经细胞。
Objective:To isolate,proliferate and differentiate neural precursor cells from the human embryonic hippocampus and to observe their morphological changes in vitro.Methods:The hippocampal cells of human abortion embryo after 8～12 weeks of gestation were cultured in DMEM/F12 with EGF,bFGF and LIF.The new-formed cellular spheres were induced to differentiate by 1% FBS and poly-L-lysine.The morphological parameters and phenotypes of those cells were measured by means of microscopy and immunocytochemical reaction.Results:In primary cultures,there were many cellular spheres,made of round cells and suspended in the medium,and some clustered or lonely cells attached to the bottom.After passages,many new spheres formed again.The cells of spheres migrated and ramified after differentiation induction,and were positive respectively in β-Ⅲ tubulin,NF-200,GFAP,Galc immunocytochemical reactions.Conclusion:The neural precursor cells derived from the human embryonic hippocampus can proliferate,divide,renew and differentiate into neuronal-limited precursors,neurons,astrocytes and oligodendrocytes in vitro.
出处
《解剖与临床》
2002年第4期134-136,F003,共4页
Anatomy and Clinics
