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目的 初步研究诱导胚胎干细胞ES D3细胞系向神经细胞定向分化可能性 ,为青光眼等视网膜视神经疾病的治疗提供可能的途径。 方法 以Buffalo大鼠肝细胞培养上清进行胚胎干细胞ES D3细胞系培养 ,参照Bain等方法进行改良 ,以视黄酸对ES D3细胞进行分化诱导 ,1d后加入阿糖胞苷 ,相差显微镜下观察细胞生长情况。 结果 加入视黄酸后 1d ,相差显微镜下可见散在 1个、2个或多个突起的神经样细胞。加入视黄酸后 2d、阿糖胞苷后第 1d ,可见大量 1条、2条或多条突起的神经样细胞 ,并相连成网络结构。 结论 视黄酸辅以阿糖胞苷在体外可成功诱导胚胎干细胞ES D3细胞系成为神经样细胞 ,并形成网络 ,结合我们裸鼠眼内接种ES D3细胞分化为神经细胞样细胞研究结果 ,胚胎干细胞定向分化及移植 ,可望成为青光眼等视网膜视神经疾病治疗的一个新的途径
OBJECTIVE: To study the possibility of inducing embryonic stem cell ES D3 cell line to differentiate into neural cells, and to provide a possible method for the treatment of retinal optic neuropathy such as glaucoma. Methods Buffalo rat hepatocyte culture supernatant was used to culture embryonic stem cells (ES D3) cell line. The method was modified by Bain et al. The differentiation of ES D3 cells was induced by retinoic acid. After 1 d, cytarabine was added and observed under a phase contrast microscope Cell growth. Results One day after the addition of retinoic acid, neurite-like cells scattered in one, two or more protuberances were observed under a phase contrast microscope. After adding retinoic acid 2 d, 1 d after cytarabine, a large number of visible 1, 2 or more neurite-like cells, and connected into a network structure. Conclusion Retinoic acid supplemented with cytarabine can successfully induce embryonic stem cell ES D3 cells to become neuroid-like cells in vitro and form a network. Combined with the results of our study on the differentiation of ES D3 cells into neuron-like cells in nude mice, Directional differentiation and transplantation of stem cells is expected to become a new approach to the treatment of retinal optic neuropathy such as glaucoma