Generation of human neural stem cell lines and brain models of hypoxic-ischemic brain damage

我们已经产生了自我更新和多能的人神经干细胞（NSCs）通过逆转录病毒介导的v-myc转移到胚胎人脑细胞，并已分离出几个克隆细胞系。这些细胞系可以保持在未分化状态，或者通过改变培养条件诱导分化。这些细胞系之一，HB 1/C4，是遗传修饰的神经干细胞，在无血清培养基中保持为稳定的细胞系，保持未定型、未分化、多能性，并表达神经干细胞特异性表型，如巢蛋白、武藏和波形蛋白。（5%胎牛血清和5%马血清），几种分化的细胞类型，其被称为NSC的后代，例如神经元，为了研究HB 1/C4细胞在髓鞘缺乏结构中的命运，我们将NB 1/C4细胞移植到成年或新生髓鞘突变Shiverer小鼠的脑中。发现HB 1/C4细胞从植入部位广泛迁移到其他解剖部位，并使以前无髓鞘的轴突重新髓鞘化。HB 1/C4细胞是一种永生化的神经干细胞，其在体外对引导信号的应答能力可引导其沿沿着多种表型途径分化，这表明HB 1/C4细胞可为人类神经系统疾病的基因细胞治疗提供一种强大的、几乎不受限制的细胞来源。

We have generated self-renewing and multipotent human neural stem cells (NSCs) via retrovirus-mediated v-myc transfer into embryonic human brain cells, and have isolated several clonal cell lines. These lines can be maintained in an undifferentiated state, or induced to differentiate by changing the culture conditions. One of these cell lines, HB1/C4, is genetically modified NSCs maintained as a stable cell line in serum-free medium remains uncommited, undifferentiated, multipotent and express phenotypes specific for NSCs such as nestin, musashi and vimentin.When NB1/C4 cells were grown in a serum containing medium (5% fetal bovine serum and 5% equine serum), several differentiated cell types, which are known as progeny of NSCs such as neurons, astrocytes and oligodendrocytes were induced in culture.To investigate the fate of HB1/C4 cells in a myelin deficiency structure, we grafted NB1/C4 cells into the brain of adult or neonatal myelin mutant shiverer mouse. HB1/C4 cells were found to migrate extensively from the site of implantation into other anatomical sites and to remyelinate formerly unmyelinated axons. The ability of the HB1/C4 cells, human immortalized NSCs, to respond in vitro to guidance cues and signals that can direct their differentiation along multiple phenotypic pathways suggests that they can providea powerful and virtually unlimitted source of cells for gene cell therapy in the human neurological diseases.