Establishment and properties of a growth factor-dependent, perpetual neural stem cell line from the human CNS
Establishment and properties of a growth factor-dependent, perpetual neural stem cell line from the human CNS
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DOI：
10.1006/exnr.1999.7237
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发表时间：
2000-01-01
影响因子：
5.3
通讯作者：
Martínez-Serrano, A
中科院分区：
文献类型：
--
作者：
Villa, A;Snyder, EY;Martínez-Serrano, A
无限量来源于人脑的神经干细胞的现成可用性对基础和应用神经科学具有极大的兴趣，包括移植后的治疗性细胞替代和基因转移。我们在这里报告的表观遗传和遗传程序的组合，使人类神经干细胞系永存。因此，我们测试了各种培养条件和基因，以获得最佳的允许人类神经干细胞连续、快速扩增和传代的条件和基因。（来源于禽逆转录病毒基因组的p110 gag-myc融合蛋白）似乎是最有效的基因;我们还确定了有丝分裂原存在的严格要求（FGF-2和EGF）在生长培养基中，实际上构成了一个有条件的永久性或永生化，一个单克隆，巢蛋白阳性，以这种方式延续的人神经干细胞系（HNSC.100）每40小时分裂一次，并在去除有丝分裂原后停止分裂，经历自发的形态分化并上调CNS中三种基本谱系（神经元、星形胶质细胞和少突胶质细胞）的标志物。因此，HNSC.100细胞保留了表观遗传扩增的人神经干细胞的基本特征。克隆分析证实了细胞系的稳定性、多能性和自我再生能力。最后，HNSC.100可以使用多种程序和编码蛋白质的基因转染和转导，用于标记目的和治疗目的（例如，人酪氨酸羟化酶I）。(C)北京大学出版社.
The ready availability of unlimited quantities of neural stem cells derived from the human brain holds great interest for basic and applied neuroscience, including therapeutic cell replacement and gene transfer following transplantation. We report here the combination of epigenetic and genetic procedures for perpetuating human neural stem cell lines. Thus we tested various culture conditions and genes for those that optimally allow for the continuous, rapid expansion and passaging of human, neural stem cells, Among them, upsilon-myc (the p110 gag-myc fusion protein derived from the avian retroviral genome) seems to be the most effective gene; we have also identified a strict requirement for the presence of mitogens (FGF-2 and EGF) in the growth medium, in effect constituting a conditional perpetuality or immortalization, A monoclonal, nestin-positive, human neural stem cell line (HNSC.100) perpetuated in this way divides every 40 h and stops dividing upon mitogen removal, undergoing spontaneous morphological differentiation and upregulating markers of the three fundamental lineages in the CNS (neurons, astrocytes, and oligodendrocytes). HNSC.100 cells therefore retain basic features of epigenetically expanded human neural stem cells, Clonal analysis confirmed the stability, multipotency, and self-renewability of the cell Line. Finally, HNSC.100 can be transfected and transduced using a variety of procedures and genes encoding proteins for marking purposes and of therapeutic interest (e,g., human tyrosine hydroxylase I). (C) 2000 Academic Press.