Enrichment of neurons and neural precursors from human embryonic stem cells
Enrichment of neurons and neural precursors from human embryonic stem cells
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DOI：
10.1006/exnr.2001.7832
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发表时间：
2001-12-01
影响因子：
5.3
通讯作者：
Rao, MS
中科院分区：
文献类型：
--
作者：
Carpenter, MK;Inokuma, MS;Rao, MS
人胚胎干细胞（hES）在体外增殖并保持其多能性超过一年（M. Amit，M. K. Carpenter，M. S.猪熊角P. Chiu，C. P.，哈里斯，M。a. Waknitz，J. Itskovitz-Eldor，and J. A.汤姆森2000. dev. 227：271 - 278），并且因此可以提供用于细胞疗法的细胞来源。在评估hES细胞分化成神经谱系的能力之前，将其在体外维持超过6个月（超过100个群体倍增）。通过形成胚状体诱导分化，随后将胚状体接种到含有有丝分裂原的成分确定的培养基中的适当基质上。这些人群中含有的细胞，巢蛋白，多唾液酸化神经细胞粘附分子（PS-NCAM）和A2B5显示阳性免疫反应性。进一步成熟后，这些细胞表达额外的神经元特异性抗原（如MAP-2，突触素和各种神经递质）。钙成像显示这些细胞对神经递质的应用有反应。电生理学分析表明，细胞膜含有电压依赖性通道，并通过电流注入触发动作电位。PS-NCAM和A2B5免疫选择或培养条件可用于产生富集的群体（60 - 90%），其可进一步分化成成熟神经元。发现hES衍生的祖细胞和神经元的性质与源自原代组织的细胞的性质相似。这些数据表明，hES细胞可以为神经祖细胞和成熟神经元提供用于治疗和毒理学用途的细胞来源。(C)2001年，爱思唯尔科学。
Human embryonic stem (hES) cells proliferate and maintain their pluripotency for over a year in vitro (M. Amit, M. K. Carpenter, M. S. Inokuma, C. P. Chiu, C. P., Harris, M. A. Waknitz, J. Itskovitz-Eldor, and J. A. Thomson. 2000. Dev. Biol. 227:271-278) and may therefore provide a cell source for cell therapies. hES cells were maintained for over 6 months in vitro (over 100 population doublings) before their ability to differentiate into the neural lineage was evaluated. Differentiation was induced by the formation of embryoid bodies that were subsequently plated onto appropriate substrates in defined medium containing mitogens. These populations contained cells that showed positive immunoreactivity to nestin, polysialylated neural cell adhesion molecule (PS-NCAM) and A2B5. After further maturation, these cells expressed additional neuron-specific antigens (such as MAP-2, synaptophysin, and various neurotransmitters). Calcium imaging demonstrated that these cells responded to neurotransmitter application. Electrophysiological analyses showed that cell membranes contained voltage-dependent channels and that action potentials were triggered by current injection. PS-NCAM and A2B5 immunoselection or culture conditions could be used to produce enriched populations (60-90%) which could be further differentiated into mature neurons. The properties of the hES-derived progenitors and neurons were found to be similar to those of cells derived from primary tissue. These data indicate that hES cells could provide a cell source for the neural progenitor cells and mature neurons for therapeutic and toxicological uses. (C) 2001 Elsevier Science.