FGF AND EGF ARE MITOGENS FOR IMMORTALIZED NEURAL PROGENITORS
FGF AND EGF ARE MITOGENS FOR IMMORTALIZED NEURAL PROGENITORS
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DOI：
10.1002/neu.480250705
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发表时间：
1994-07-01
期刊：
影响因子：
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通讯作者：
GOTTLIEB, DI
中科院分区：
文献类型：
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作者：
KITCHENS, DL;SNYDER, EY;GOTTLIEB, DI
来自新生小鼠小脑外胚层的单个神经祖细胞，先前通过逆转录病毒介导的禽myc（v-myc）转导而永生化。C17-2是那些克隆多能祖细胞系之一（Snyder等人，1992，Cell 68：33-51; Ryder等人，1990，J. Neurobiol. 21：356-375）。当移植到新生小鼠小脑（CB）中时，细胞参与正常的CB发育;它们以细胞结构适当、非致瘤的方式移植，并分化成与内源性祖细胞类似的多种CB细胞类型（神经元和神经胶质）（Snyder等人）。，1992年，同上）。它们似乎还沿着神经轴和在多个其他阶段植入并参与多个其他结构的发育（Snyder等人，1993年，SEC. Neurosci.摘要19）。因此，关于这些永生化祖细胞的结论可能适用于体内内源性神经祖细胞。为了帮助鉴定和分析促进内源性祖细胞分化的因素，我们首先研究了在确定的无血清培养基（N2）中维持C17-2细胞的能力。细胞在体外N2中存活，但以非常低的速率进行有丝分裂。然而，来自小鼠颌下腺或人重组蛋白的表皮生长因子（EGF）的加入明显刺激胸苷掺入和细胞分裂约三倍。碱性成纤维细胞生长因子（bFGF）是一种更有效的促有丝分裂原，促进胸苷掺入、细胞分裂和细胞数量的净增加，与血清中的细胞数量相等。EGF和bFGF在非常低的纳摩尔浓度下都有活性，这表明它们与各自的受体相互作用，而不是同源受体系统。研究结果表明，C17-2细胞可以在完全限定的培养基中维持和增殖，为分析其他生长和分化因子提供了基础。EGF和特别是bFGF对这些细胞是促有丝分裂的，这与最近对原代神经组织的观察结果雅阁（Reynolds和韦斯，1992，Science 255：1707-1710; Kilpatrick和Bartlett，1993，Neuron 10：255-265; Ray等人，1993，Proc.Natl. Acad. Sci. USA 90：3602-3606），表明bFGF和EGF反应性可能是神经祖细胞的基本性质。(C)1994年，John Wiley and Sons，Inc.
Individual neural progenitors, derived from the external germinal layer of neonatal murine cerebellum, were previously immortalized by the retrovirus-mediated transduction of avian myc (v-myc). C17-2 is one of those clonal multipotent progenitor cell lines (Snyder et al., 1992, Cell 68: 33-51; Ryder et al., 1990, J. Neurobiol. 21:356-375). When transplanted into newborn mouse cerebellum (CB), the cells participate in normal CB development; they engraft in a cytoarchitecturally appropriate, nontumorigenic manner and differentiate into multiple CB cell types(neuronal and glial) similar to endogenous progenitors (Snyder et al., 1992, as above). They also appear to engraft and participate in the development of multiple other structures along the neural axis and at multiple other stages (Snyder et al., 1993, Sec. Neurosci. Abstr. 19). Thus conclusions regarding these immortalized progenitors may be applicable to endogenous neural progenitors in vivo. To help identify and analyze factors that promote differentiation of endogenous progenitors, we first investigated the ability to maintain C17-2 cells in a defined, serum-free medium (N2). The cells survive in vitro in N2 but undergo mitosis at a very low rate, Addition of epidermal growth factor (EGF), however, either from mouse submaxillary gland or the human recombinant protein, appreciably stimulates thymidine incorporation and cell division approximately threefold. Basic fibroblast growth factor (bFGF) is an even more potent mitogen, promoting thymidine incorporation, cell division, and a net increase in cell number equal to that in serum. Both EGF and bFGF are active at very low nanomolar concentrations, suggesting that they interact with their respective receptors rather than a homologous receptor system. The findings demonstrate that C17-2 cells can be maintained and propagated in a fully defined medium, providing the basis for analysis of other growth and differentiation factors. That EGF and particularly bFGF are mitogenic for these cells is in accord with recent observations on primary neural tissue (Reynolds and Weiss, 1992, Science 255:1707-1710; Kilpatrick and Bartlett, 1993, Neuron 10:255-265; Ray et al., 1993, Proc. Natl. Acad. Sci. USA 90: 3602-3606) suggesting that bFGF and EGF responsiveness may be fundamental properties of neural progenitors. (C) 1994 John Wiley and Sons, Inc.