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ISOLATION AND EXPANSION OF HUMAN EMBRYONIC NEURAL STEM/PROGENITOR CELLS IN VITRO

ISOLATION AND EXPANSION OF HUMAN EMBRYONIC NEURAL STEM/PROGENITOR CELLS IN VITRO
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摘要 Objective:To isolate, culture and identify human embryonic neural stem cells and to establish a practical passaging method.Method:The cerebral cortex cells were isolated from aborted embryos (11~13 weeks) by mechanical dissociation,and cultured in DMEM/F12 culture medium supplemented with N2 and growth factors for proliferation. Upon passaging, the neurospheres were pipetted gentlely to separate them into several cell masses and then grown in growth medium. The cells were grown in DMEM/F12 medium with serum (without growth factors) to induce differentiation. The stem cell, neuron, astrocyte and oligodendrocyte were identified by immunocytochemistry with antibodies to vimentin, MAP 2, GFAP and GalC, respectively. Results:The primary cells grew together and formed neurospheres at 5 th ~7 th day. They were all vimentin positive and could be passaged for at least 8 passages. After passaging, the cell masses grew up and formed new neurospheres rapidly.These cells could differentiated into MAP 2(+),GFAP(+) or GalC(+) cells.Conclusion:The neural stem cells from human embryonic cerebral cortex have the capacity of proliferation and multi-differentiation in vitro. The passaging methods we used in this experiment were practical and convenient. Objective:To isolate, culture and identify human embryonic neural stem cells and to establish a practical passaging method.Method:The cerebral cortex cells were isolated from aborted embryos (11~13 weeks) by mechanical dissociation,and cultured in DMEM/F12 culture medium supplemented with N2 and growth factors for proliferation. Upon passaging, the neurospheres were pipetted gentlely to separate them into several cell masses and then grown in growth medium. The cells were grown in DMEM/F12 medium with serum (without growth factors) to induce differentiation. The stem cell, neuron, astrocyte and oligodendrocyte were identified by immunocytochemistry with antibodies to vimentin, MAP 2, GFAP and GalC, respectively. Results:The primary cells grew together and formed neurospheres at 5 th ~7 th day. They were all vimentin positive and could be passaged for at least 8 passages. After passaging, the cell masses grew up and formed new neurospheres rapidly.These cells could differentiated into MAP 2(+),GFAP(+) or GalC(+) cells.Conclusion:The neural stem cells from human embryonic cerebral cortex have the capacity of proliferation and multi-differentiation in vitro. The passaging methods we used in this experiment were practical and convenient.
出处 《中国现代医学杂志》 CAS CSCD 2002年第23期15-19,共5页 China Journal of Modern Medicine
基金 ThisworkwassupportedbytheNationalNaturalScienceFoundation(No .3 0 170 3 0 0 ) NaturalScienceFoundationofShanxiProvince(2 0 0 1SM63 )andFoundationofsecondhospitalofxi’anjiaotonguniversity(2 0 0 0KG -18)
关键词 人胚神经 干细胞/祖细胞 分离 体外增殖 Neural Stem/Progenitor Cell Isolation and Expansion Human Cortex
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参考文献12

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