起源于小鼠诱导性多能干细胞的大脑皮质类器官的建立及其生物学特性

doi:10.16098/j.issn.0529-1356.2017.04.004

解剖学报 ›› 2017, Vol. 48 ›› Issue (4): 387-396.doi: 10.16098/j.issn.0529-1356.2017.04.004

起源于小鼠诱导性多能干细胞的大脑皮质类器官的建立及其生物学特性

范文娟王倩孙仪征王来邓锦波*

河南大学生命科学学院,河南大学神经生物学研究所，河南开封 475004

收稿日期:2016-08-29 修回日期:2016-11-08 出版日期:2017-08-06 发布日期:2017-08-06
通讯作者: 邓锦波 E-mail:jinbo_deng@henu.edu.cn
基金资助:
河南大学省属高校科研基金;河南大学省属高校科研基金;河南省自然科学基金;河南省自然科学基金

Mouse induced pluripotent stem cell-derived cortical organoids and its biological characteristics

FAN Wen-juan WANG Qian SUN Yi-zheng WANG Lai DENG Jin-bo*

Institute of Neurobiology, School of Life Science, He’nan University, He’nan Kaifeng 475004, China

Received:2016-08-29 Revised:2016-11-08 Online:2017-08-06 Published:2017-08-06
Contact: DENG Jin-bo E-mail:jinbo_deng@henu.edu.cn
Supported by:
Henan University Youth Training Foundation;Henan University Youth Training Foundation

摘要/Abstract

摘要：

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。

关键词: 诱导性多能干细胞, 大脑皮质类器官, 三维细胞培养, 神经再生, 干细胞移植, 免疫荧光, 小鼠

Abstract:

Objective To culture cerebral organoid from mouse induced pluripotent stem cells (iPSCs) and analyze its biological characteristics. Methods iPSCs were cultured into embryoid bodies (EB) through suspension cultures. EB were induced with some nervous growth factors and transfered to Matrigel droplet. After 1-2 months culture, cerebral organoids were formed. Immunolabeling, transmission electron microscopy, slice co-culture and DiO tracing were carried out to identify their biological characteristics. Results iPSCs-derived cerebral cortical organoids differentiated into neural precursor cells, nervous cells and glial cells. The cerebral organoid had cortical lamination, contained neuroepithelium, cortical plate and molecular layer and had the ability of neural regeneration and neural repair. Conclusion The cerebral organoid derived from mouse iPSCs l was cultured successfully, and the cultured cerebral organoid had similar biological characteristics with mammal cerebrum, including neural differentiation, cortical lamination, and especially neural regeneration and repair.

Key words: Induced pluripotent stem cells cells, Cerebral cortical organoid, Three dimensional culture, Neuroregeneration, Stem cell transplantation, Immunofluorescence, Mouse

范文娟王倩孙仪征王来邓锦波. 起源于小鼠诱导性多能干细胞的大脑皮质类器官的建立及其生物学特性[J]. 解剖学报, 2017, 48(4): 387-396.

FAN Wen-juan WANG Qian SUN Yi-zheng WANG Lai DENG Jin-bo. Mouse induced pluripotent stem cell-derived cortical organoids and its biological characteristics[J]. Acta Anatomica Sinica, 2017, 48(4): 387-396.

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起源于小鼠诱导性多能干细胞的大脑皮质类器官的建立及其生物学特性

Mouse induced pluripotent stem cell-derived cortical organoids and its biological characteristics

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