肝细胞生长因子促进猕猴胚胎干细胞来源的神经前体细胞的增殖

Hepatocyte Growth Factor Promotes the Proliferation of the Neural Progenitors Derived from Rhesus Monkey Embryonic Stem Cells

肝细胞生长因子(hepatocyte growth factor,HGF)是一个多效应因子,在神经系统中具有重要作用。早前的研究发现采用HGF和G5 supplement结合EB(embryoid body)法可诱导猕猴胚胎干细胞(rhesus embryonic stem cells,rESCs)定向分化成高纯度的可移植的神经前体细胞(neural progenitors),但对于HGF在整个诱导分化过程中的具体作用及机制还不清楚。本研究改进先前研究体系,采用单层培养法,同时添加HGF和bFGF(basic fibroblast growth factor,碱性成纤维细胞生长因子)诱导rESCs在两周内定向分化为高纯度[(81.66±4.37)%]的神经前体细胞,并且单独添加HGF或bFGF以及两者都没有添加的条件下也得到了相似比例的神经前体细胞,表明外源性的HGF在诱导rESCs向神经前体细胞转变的过程中对于神经细胞命运的决定并不起作用;进一步研究发现HGF能有效地促进神经前体细胞的增殖,并且与bFGF具有协同作用。总之,本研究建立了一种更为简单的诱导rESCs分化成神经细胞的方法,发现外源性的HGF在rESCs向神经前体细胞分化的过程中并没有神经诱导的作用,但能与bFGF协同作用促进rESCs来源的神经前体细胞的增殖。

Hepatocyte growth factor （HGF） is a pleiotropic factor, which plays an important role in the nervous system. Previously, we had produced highly enriched （88.3 ±8.1%） transplantable populations of neural progenitors from rhesus monkey embryonic stem cells （rESCs） using an embryoid body （EB） system containing HGF and G5 supplement. However, the exact role of HGF in the neural differentiation process as well as the mechanism of HGF is not clear. Here, on the basis of the ameliorated differentiation system, we efficiently produced populations of neural progenitors （81.66 ± 4.37%） from rESCs in an adherent monoculture system containing HGF and basic fibroblast growth factor （bFGF） within two weeks. Moreover, we obtained similar results with the systems containing either HGF or bFGF alone, or neither of them, which implied that extrinsic HGF can not determine the neural fate of the rESCs in the differentiation process. Further results suggested HGF with bFGF synergistically promoted the proliferation of the neural progenitors derived from rESCs. In conclusion, through this study, a much simpler method of inducing rESCs differentiation towards neural cells has been established and extrinsic HGF did not function in neural induction from rESCs to neural progenitor cells. However, HGF with bFGF synergistically promoted the proliferation of the neural progenitors derived from rESCs.