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目的探讨人胚胎纹状体区神经干细胞在体外的生物学特性。方法从16~20周人胚胎纹状体区分离培养神经干细胞,在体外进行传代、分化,应用免疫组织化学荧光染色等方法,对此区神经干细胞在体外增殖和分化等情况进行研究。结果纹状体区神经干细胞在体外原代培养扩增1月内分裂增殖速度快,在含有EGF和bFGF的培养基中生长最为良好,平均倍增时间为3~4 d。分化培养后可以形成各种神经细胞,在原代培养后2周分化情况好,分化为神经元比例高,约占50%左右,在原代培养8周以后分化为神经元的比例下降,约占20%左右。结论人胚胎纹状体区的神经干细胞在体外有较强的自我更新和增殖能力,并表达了干细胞的原始特征,在体外传代过程中,增殖速度随着时间不断下降,其分化为各种神经细胞的能力也不同。培养基中的有丝分裂原对于神经干细胞的增殖和分裂的影响不同。神经球并非均质的,内部仅有部分细胞在分裂增殖。
Objective To investigate the biological characteristics of neural stem cells in human embryonic striatum in vitro. Methods Neural stem cells (NSCs) were isolated and cultured from human embryonic striatum in 16 ~ 20 weeks and passaged and differentiated in vitro. The proliferation and differentiation of neural stem cells in vitro were studied by immunohistochemical staining. Results The stromal cells proliferated rapidly in primary cultured and proliferated cells in vitro in the striatum. The strongest growth was found in media containing EGF and bFGF with average doubling time of 3-4 days. Differentiation and culture can form a variety of nerve cells, 2 weeks after the primary culture well differentiated into high proportion of neurons, accounting for about 50%, 8 weeks after the primary culture differentiated into neurons decreased, accounting for about 20 %about. Conclusion The neural stem cells in human embryonic striatum have strong ability of self-renewal and proliferation in vitro and express the original characteristics of stem cells. In the process of passage in vitro, the proliferation rate of neural stem cells declines with the time and differentiates into various nerves The ability of cells is also different. Mitochondria in culture have different effects on the proliferation and division of neural stem cells. Neurospheres are not homogeneous, only part of the internal cell proliferation in the division.