Generation of neural stem cells from discarded human fetal cortical tissue.
Generation of neural stem cells from discarded human fetal cortical tissue.
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DOI：
10.3791/2681
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发表时间：
2011-05-25
期刊：
影响因子：
--
通讯作者：
Sheen, Volney L
中科院分区：
文献类型：
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作者：
Lu, Jie;Delli-Bovi, Laurent C;Sheen, Volney L
神经干细胞（NSCs）在皮层板发育过程中沿脑室带神经上皮生长。这些早期祖细胞最终形成中间祖细胞，然后形成形成大脑皮层的各种神经元和胶质细胞亚型。从废弃的正常胎儿组织中产生和扩展人类神经干细胞（所谓的神经球）的能力为直接研究正常人类神经干细胞发育的功能方面提供了一种手段。这种方法也可以用于从已知的神经系统疾病中产生NSCs，从而为识别改变祖细胞增殖、迁移和分化的疾病过程提供机会。我们专注于确定人类唐氏综合征NSCs可能加速阿尔茨海默病表型的病理机制。无论是体内还是体外小鼠模型都不能复制位于人类21号染色体上的相同基因库。本研究采用一种简单可靠的方法从流产的人类胎儿皮质中分离唐氏综合征NSCs并进行培养。该方法提供了采集组织的具体方面，具有有限解剖标志的解剖，细胞分选，人类NSCs的镀层和传代。我们还提供了一些诱导人类NSCs分化为更有选择性的细胞亚型的基本方案。
Neural stem cells (NSCs) reside along the ventricular zone neuroepithelium during the development of the cortical plate. These early progenitors ultimately give rise to intermediate progenitors and later, the various neuronal and glial cell subtypes that form the cerebral cortex. The capacity to generate and expand human NSCs (so called neurospheres) from discarded normal fetal tissue provides a means with which to directly study the functional aspects of normal human NSC development. This approach can also be directed toward the generation of NSCs from known neurological disorders, thereby affording the opportunity to identify disease processes that alter progenitor proliferation, migration and differentiation. We have focused on identifying pathological mechanisms in human Down syndrome NSCs that might contribute to the accelerated Alzheimer's disease phenotype. Neither in vivo nor in vitro mouse models can replicate the identical repertoire of genes located on human chromosome 21. Here we use a simple and reliable method to isolate Down syndrome NSCs from aborted human fetal cortices and grow them in culture. The methodology provides specific aspects of harvesting the tissue, dissection with limited anatomical landmarks, cell sorting, plating and passaging of human NSCs. We also provide some basic protocols for inducing differentiation of human NSCs into more selective cell subtypes.