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小鼠胚胎干细胞体外分化为神经前体细胞的研究
引用本文:吴旋,黎海蒂,李树浓,徐海伟,徐令.小鼠胚胎干细胞体外分化为神经前体细胞的研究[J].第三军医大学学报,2001,23(11):1260-1263.
作者姓名:吴旋  黎海蒂  李树浓  徐海伟  徐令
作者单位:1. 第三军医大学基础医学部生理学教研室,
2. 中山医科大学病理生理学教研室
3. Department of Medicine Beth Israel Deaconess Hospital Harvard Medical School USA
基金项目:国家自然科学基金资助项目 ( 3990 0 169,30 0 70 72 7)
摘    要:目的 探索小鼠胚胎干细胞(Embryonic stem cell,ES cell)体外分化为神经前体细胞(Neural precursor cells,NPC)的无血清培养条件,比较人胚胎成纤维细胞(Human embryonic fibroblasts,HEF)与小鼠胚胎成纤维细胞(Mouse embryonic fibroblasts,MEF)对小鼠ES细胞生长的作用。方法 在MEF或HEF饲养层上培养ES细胞,培养液中含白血病抑制因子。采用无血清方法培养NPC,免疫组化方法检测巢蛋白(Nestin),用硝基四氮啖蓝/5-溴-4-氯-吲哚基磷酸(NBT/BCIP)显色检测碱性磷酸酶。结果 无血清培养可以获得86%的NPC。HEF与MEF-样能维持ES未分化状态。结论 无血清培养方法有利于ES细胞向NPC分化,HEF可用于小鼠ES细胞的培养,而且比MEF优越。

关 键 词:胚胎干细胞  神经前体细胞  无血清培养  人胚胎成纤维细胞  小鼠
文章编号:1000-5404(2001)11-1260-04
修稿时间:2001年2月20日

Differentiation of mouse embryonic stem cells to neural precursor cells in vitro
WU Xuan,LI Hai di,LI Shu nong,XU Hai wei,XU Lin.Differentiation of mouse embryonic stem cells to neural precursor cells in vitro[J].Acta Academiae Medicinae Militaris Tertiae,2001,23(11):1260-1263.
Authors:WU Xuan  LI Hai di  LI Shu nong  XU Hai wei  XU Lin
Abstract:Objective To explore the serum free culture condition of the differentiation of embryonic stem cells(ES) to neural precursor cells (NPC) and compare the effects of human embryonic fibroblasts (HEF) as the feeder layer of ES with that of mouse embryonic fibroblasts (MEF). Methods Mouse embryonic stem cells were cultured on the feeder layer of HEF or MEF in the medium containing leukemia inhibitory factor to inhibit differentiation. NPC were selectively cultured in serum free medium. Nestin antigen was detected with immunocytochemical methods and alkaline phosphatase activity was visualized with NBT/BCIP. Results About 86% NPC were obtained from ES cells in the serum free culture condition. HEF could be used as feeder layer to culture mouse ES cells and even better than MEF. Conclusion Results show that NPC of high purity could be cultured from ES cells with serum free culture method. HEF can replace MEF and with more benefits.
Keywords:embryonic stem cell  neural precursor cell  serum  free culture  human embryonic fibroblast
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