不同促细胞分裂因子对人神经干细胞定向分化的影响

Effect of mitosis-promoting factors on regulation of human NSCs orientating differentiation

目的观察不同促细胞分裂因子对人神经干细胞(NSCs)增殖及定向分化的影响。方法对人NSCs用无血清DMEM培养基行原代培养的同时,分别加入表皮生长因子(EGF)、碱性成纤维生长因子(bFGF)、神经生长因子(NGF)、维甲酸(RA)等因子,观察其对NSCs定向分化的作用。结果EGF培养的NSCs,克隆球形成慢且较松散,经血清诱导分化后,主要为星形胶质细胞,仅有少数神经元。而bFGF培养的NSCs则生长良好。在加血清诱导分化后,不同浓度bFGF培养的NSCs分化的细胞不同。bFGF与EGF共同培养的NSCs,其生长及神经球形成良好。在加血清诱导分化后,分化的神经细胞比例更接近脑内神经细胞的组分。NGF对神经球的形成无明显影响,但可促使其向神经元分化。RA使神经克隆球形成,可使NSCs直接分化为神经元细胞的比例明显增加。结论不同促细胞分裂因子对人NSCs的增殖及定向分化均有一定影响。

Objective To explore the effect of mitasis-pmmoting factors （MPF） on regulation of neural stem cells （NSCs） proliferation and orientating differentiation. Methods First, we cultured the NSCs from hippocanpus of 10 to 14 w embryo brains, then incubated the NSCs in different free-serum-DMEM medium containing human epidermal growth factor （EGF）, basic fibmblast growth factor （bFGF）, retinoic acid （RA） and nerve growth factor （NGF） respectively. The effect of these factors on orientating differentiation of NSCs was observed by immunocytochemical staining methods. Results The neurospheres formed slowly and were incompact in EGF group. NSCs could be induced to differentiate mostly into glial cells and only a few into neurons in tiffs group. The NSCs grew very well in the medium containing bFGF. Both the neurospheres and NSCs developed well in the medium containing both bFGE and EGF. The NSCs could be induced to differentiate into glial cells and neurons and the ratio of them was similar to that in vivo, when NSCs were cultured in DMEM medium supplemented with fetal bovine serum in the bFGE combined EGF group. NGF had no significant influence on the proliferation of NSCs, however, it can promote the differentiation of NSCs into the neurons. RA could promote the formation of neurospheres, and could increase the differentiation rate of NSCs into neurons. Conclusion Different mitasis-pmmoting factors have some kinds of ability to affect the proliferation and orientating differentiation of NSCs.