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人类胚胎干细胞是替代治疗充满希望的细胞来源.描述了从人胚胎干细胞诱导分化出神经干细胞的方法.将人胚胎干细胞系PKU1,PKU2在细菌培养皿中悬浮培养,分化形成囊性拟胚体.拟胚体接种至组织培养皿,加入N2培养液和生长因子bFGF培养2周,拟胚体贴壁、展开,中心出现灶状增生,有突起的小细胞.用机械方法取下此种细胞,重新接种,则细胞团悬浮生长,形成神经球.培养10天后,将神经球打散成单细胞接种,该细胞贴壁生长旺盛.免疫荧光检测显示为几乎100%纯净的nestin阳性细胞.将培养液中的生长因子撤除,继续培养7~10天,细胞分化为神经元,该细胞呈现β-tubulinisotypeⅢ阳性、GABA阳性、serotonin阳性、synaptophysin阳性.在生长因子PDGF-AA诱导下,细胞分化为星形胶质细胞,其GFAP阳性;或少突胶质细胞,其O4阳性.可见,人类胚胎干细胞经上述方法培养可分化为典型神经干细胞,表达神经干细胞特异的标志分子nestin、能自我更新、具有分化为神经系统三类主要细胞的能力.
Human embryonic stem cells (NSCs) are a promising source of alternative therapies and describe a method of differentiating neural stem cells from human embryonic stem cells (NSCs). Human embryonic stem cell lines, PKU1 and PKU2, are suspended in bacterial culture dishes and differentiated to form cystic embryoid bodies. The embryoid bodies were inoculated into tissue culture dishes and cultured in N2 medium and growth factor bFGF for 2 weeks. The embryoid bodies were adherent and expanded, with focus-like hyperplasia and prominent small cells in the center. These cells were removed mechanically and re- After inoculation, the cell pellets were suspended and grown to form neurospheres, and after 10 days of culture, the neurospheres were inoculated into a single cell inoculation and the cells grew well on the wall, and immunofluorescence assay showed almost 100% pure nestin positive cells. The cells were differentiated into neurons, showing β-tubulinisotype Ⅲ positive, GABA positive, serotonin positive and synaptophysin positive. Under the induction of growth factor PDGF-AA, the cells differentiated into star Glial cells, the GFAP positive; or oligodendrocytes, O4 positive.It can be seen, human embryonic stem cells can be cultured by the above method can be differentiated into typical neural stem cells, the expression of God Stem cell-specific markers Nestin molecules, self-renewing, the ability to differentiate into three main nervous system cells.