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目的利用小鼠诱导性多能干细胞(iPSCs)建立大脑皮质类器官并分析其生物学特性。方法首先将生长在饲养层细胞上的iPSCs克隆消化成单细胞,悬浮培养使其自发形成拟胚体。随后,对其进行神经上皮分化的诱导。在合适时间将其嵌入Matrigel基质胶做的悬滴中,摇动培养,形成大脑皮质类器官;最后利用免疫荧光、Dio示踪、透射电子显微镜等技术检测大脑皮质类器官的生物学特性。结果大脑皮质类器官有神经上皮、神经元及神经胶质细胞的分化特征;具有简单的片层化结构,如皮质板和分子层,且大脑皮质类器官具有神经再生及修复能力。结论利用iPSCs成功建立了大脑皮质类器官模型,且该模型具有与活体大脑皮质相似的特征,且具有神经再生能力。
Objective To establish cerebral cortical organs using mouse induced pluripotent stem cells (iPSCs) and analyze their biological characteristics. Methods Firstly, iPSCs growing on feeder cells were cloned and digested into single cells. The cells were cultured in suspension to form spontaneous embryoid bodies. Subsequently, it is induced for neural epithelial differentiation. At the right time, it was embedded in the hanging drop made of Matrigel matrices and shaken to form cerebral cortex organ. Finally, the biological characteristics of cerebral cortex organ were detected by immunofluorescence, Dio tracing and transmission electron microscopy. Results The cerebral cortical organs had the differentiation characteristics of neuroepithelial, neuronal and glial cells. They had simple lamellar structures such as cortical plate and molecular layer, and the cortical organs of the cerebral cortex had nerve regeneration and repair ability. Conclusion The model of cerebral cortical organ was successfully established by using iPSCs. The model has similar characteristics with living cerebral cortex and has the ability of regenerating nerve.