Yang Tao, Tsang Kam Sze, Poon Wai Sang, Ng Ho Keung
Li Ka Shing Institute of Health Sciences, The Chinese University of Hong Kong, Hong Kong; Department of Anatomical and Cellular Pathology, The Chinese University of Hong Kong, Prince of Wales Hospital, Hong Kong.
Cell Transplant. 2009;18(4):391-404. doi: 10.3727/096368909788809767.
Embryonic stem (ES) cell-derived cell products may serve as a source of cells for regenerative medicine. Currently available technologies for the induction of ES cells into neural lineage cells require extended culturing in vitro and complex procedural manipulations, with variable yields of heterogeneous cells, which have hindered the prospective use of cell derivatives for treatment of ischemic stroke. We established a simple and efficient method to derive mouse ES cells into neural lineage cells using an 8-day coculture with the bone marrow stromal cells MS5, followed by a 6-day propagation culture and a 4-day selection culture. The protocol generated a relatively high yield of neural lineage cells without any mesodermal and endodermal lineage commitment. In in vivo study, these derived cells could improve the cognitive function of ischemic stroke mice. Three weeks after transplantation, migration of implanted cells to lesioned areas was noted. It was also evident of a normalization of pyramidal neuron density and morphology in hippocampal CA1 region. One (1/17) episode of teratoma development was noted. Data suggested that MS5 cells may exert a neurotrophic effect to enhance neural differentiation of ES cells and MS5-induced ES cell-derived cells appeared to be applicable to cell therapy for ischemic stroke.
胚胎干细胞（ES细胞）衍生的细胞产物可作为再生医学的细胞来源。目前将ES细胞诱导为神经谱系细胞的可用技术需要在体外进行长时间培养和复杂的程序操作，产生的异质细胞产量各不相同，这阻碍了细胞衍生物在缺血性中风治疗中的预期应用。我们建立了一种简单有效的方法，将小鼠ES细胞与骨髓基质细胞MS5共培养8天，然后进行6天的增殖培养和4天的选择培养，从而将其诱导为神经谱系细胞。该方案产生了相对较高产量的神经谱系细胞，且没有任何中胚层和内胚层谱系的分化。在体内研究中，这些衍生细胞可以改善缺血性中风小鼠的认知功能。移植三周后，观察到植入细胞向损伤区域迁移。海马CA1区锥体细胞密度和形态的正常化也很明显。记录到1例（1/17）畸胎瘤发生。数据表明，MS5细胞可能发挥神经营养作用，增强ES细胞的神经分化，并且MS5诱导的ES细胞衍生细胞似乎适用于缺血性中风的细胞治疗。
