Lie Khun-Hong, Chung Henry C Y, Sidhu Kuldip S
Stem Cell Laboratory, Faculty of Medicine, School of Psychiatry, University of New South Wales, Randwick, NSW, Australia.
Methods Mol Biol. 2012;873:237-46. doi: 10.1007/978-1-61779-794-1_15.
The differentiation of human embryonic stem cells (hESCs) and human-induced pluripotent stem cells (hiPSCs) towards functional neurons particularly hold great potential for the cell-based replacement therapy in neurodegenerative diseases. Here, we describe a stepwise differentiation protocol that mimics the early stage of neural development in human to promote the generation of neuroprogenitors at a high yield. Both the hESCs and hiPSCs are initially cultured in an optimized feeder-free condition, which offer an efficient formation of aggregates. To specify the neuroectodermal specification, these aggregates are differentiated in a defined neural induction medium to develop into neural rosettes-like structures. The rosettes are expanded into free-floating sphere and can be further propagated or developed into variety of neuronal subtypes.
人类胚胎干细胞（hESCs）和人类诱导多能干细胞（hiPSCs）向功能性神经元的分化对于神经退行性疾病的细胞替代疗法具有巨大潜力。在此，我们描述了一种逐步分化方案，该方案模拟人类神经发育的早期阶段，以高产率促进神经祖细胞的生成。hESCs和hiPSCs最初都在优化的无饲养层条件下培养，这有利于高效形成聚集体。为了确定神经外胚层的特化，这些聚集体在特定的神经诱导培养基中分化，发育成神经玫瑰花结样结构。玫瑰花结被扩增成自由漂浮的球体，并可进一步增殖或发育成多种神经元亚型。
