Cho Myung-Soo, Hwang Dong-Youn, Kim Dong-Wook
Stem Cell Research Center, 21C Frontier R&D Program of Ministry of Education, Science and Technology, Yonsei University Medical Centre, 134 Sinchon-dong, Seodaemun-gu, Seoul 120-752, Korea.
Nat Protoc. 2008;3(12):1888-94. doi: 10.1038/nprot.2008.188.
Cell-replacement therapy using human embryonic stem cells (hESCs) holds great promise in treating Parkinson's disease. We have recently reported a highly efficient method to generate functional dopaminergic (DA) neurons from hESCs. Our method includes a unique step, the formation of spherical neural masses (SNMs), and offers the highest yield of DA neurons ever achieved so far. In this report, we describe our method step by step, covering not only how to differentiate hESCs into DA neurons at a high yield, but also how to amplify, freeze and thaw the SNMs, which are the key structures that make our protocol unique and advantageous. Although the whole process of generation of DA neurons from hESCs takes about 2 months, only 14 d are needed to derive DA neurons from the SNMs.
使用人类胚胎干细胞（hESC）进行细胞替代疗法在治疗帕金森病方面具有巨大潜力。我们最近报道了一种从hESC高效生成功能性多巴胺能（DA）神经元的方法。我们的方法包括一个独特步骤，即形成球形神经团块（SNM），并且实现了迄今为止最高的DA神经元产量。在本报告中，我们逐步描述我们的方法，不仅涵盖如何以高产率将hESC分化为DA神经元，还包括如何扩增、冷冻和解冻SNM，这些关键结构使我们的方案独具特色且具有优势。尽管从hESC生成DA神经元的整个过程大约需要2个月，但从SNM衍生出DA神经元仅需14天。
