Yin Dezhong, Tavakoli Tahereh, Gao Wei-Qiang, Ma Wu
Stem Cell Center, American Type Culture Collection, Manassas, VA, USA.
Methods Mol Biol. 2012;873:247-59. doi: 10.1007/978-1-61779-794-1_16.
Neural differentiation of human embryonic (ES) and induced pluripotent (iPS) stem cell lines has been used for research in early human development, drug discovery, and cell replacement therapies. It is critical to establish generic differentiation protocols to compare the neural specification potential of each individually derived pluripotent stem cell line and identify the efficacious lines for research and therapeutic use. Here, we describe a reproducible and quantitative protocol to assess the neural progenitor (NP) generation of human pluripotent stem cell lines. This method includes a robust and well-defined neural inducing platform for Pax6(+) neural rosette (neuroectodermal cells) generation, propagation, and subsequent differentiation into nestin(+) NPs. A side-by-side comparison under common culture conditions among three human ES cell lines, TE03, TE06, and BG01V, and one iPS cell line, HD02, showed highly variable efficiency in their differentiation into NPs.
人类胚胎干细胞(ES)系和诱导多能干细胞(iPS)系的神经分化已被用于早期人类发育研究、药物发现和细胞替代疗法。建立通用的分化方案对于比较每个单独衍生的多能干细胞系的神经定向潜能,并确定用于研究和治疗用途的有效细胞系至关重要。在此,我们描述了一种可重复且定量的方案,用于评估人类多能干细胞系的神经祖细胞(NP)生成情况。该方法包括一个强大且定义明确的神经诱导平台,用于生成、增殖Pax6(+)神经玫瑰花结(神经外胚层细胞),并随后分化为巢蛋白(+)NP。在共同培养条件下对三个人类ES细胞系TE03、TE06和BG01V以及一个iPS细胞系HD02进行的并行比较显示,它们分化为NP的效率差异很大。