摘要
目的 探讨体外培养大脑类器官与体内正常大脑皮质发育的差异。方法 1.分组:在体大脑皮质组、体外培养大脑皮质类器官组。2.取材:收取昆明小鼠胚胎7.5 d(E7.5)、E9.5完整胚胎以及E11.5、E14.5、出生后3 d(P3)、P7大脑组织,每组标本取3只。利用小鼠诱导性多能干细胞(iPSCs)培养大脑皮质类器官,收取不同培养时间点的标本,每时间点取3个以上。3.检测方法:通过免疫荧光染色分析各组标本不同类型细胞的分布。结果 大脑类器官在组织发生、神经细胞与神经胶质细胞分化等方面与活体大脑皮质发育类似,但大脑类器官尚不具有活体大脑皮质复杂的片层结构。结论 大脑类器官可以模拟在体大脑的发育,用于疾病模型建立,但大脑类器官的培养技术仍需改进。
Objective To understand the characteristics and developmental differences between cerebral organoidsin vitroand normal cerebral corticesin vivo.Methods 1.Grouping:cerebral corticesin vivogroup and cultured cerebralorganoidsin vitrogroup.2.Sample collection:cortical tissues were collected from Kunming mouse embryos at embryonicday 7.5(E7.5),E9.5,E11.5,E14.5,and postnatal day 3(P3)or P7.Three specimens were taken from each group.Meanwhile,cerebral organoids were cultured with mouse induced pluripotent stem cells(iPSCs),and samples at differentculture time point were collected,and more than 3 samples were collected at each time point.3.Detection method:thedistribution of different types of cells in each group of specimens was analyzed by immunofluorescent staining.ResultsWhile relative similarities betweenin vivocerebral cortical development and the cerebral organoidsin vitrowere observed,including the histogenesis,and the morphological differentiation of nerve cells and glial cells,the lamellar architecture ofcerebral cortex in mouse brain was not observed in cerebral organoids.Conclusion The development of cerebral organoidsin vitrohas some similarity with body’s cortical development.Therefore,cerebral organoids can be used to a substitution ofcortex and diseases’models,but improvement of the existing technologies is necessary.
作者
范文娟
陈旭东
陈永芳
杨旭光
金少举
赵志军
邓锦波
FAN Wen-juan;CHEN Xu-dong;CHEN Yong-fang;YANG Xu-guang;JIN Shao-ju;ZHAO Zhi-jun;DENG Jin-bo(Luohe Medical College,He’nan Province Engineering Research Center of Nutrition and Health,He’nan Luohe462002,China;Neurology Department,the First People’s Hospital of Luohe City,He’nan Luohe462000,China)
出处
《解剖学报》
CAS
CSCD
北大核心
2023年第4期383-391,共9页
Acta Anatomica Sinica
基金
河南省科技厅项目(212102310891,212102310896)
河南省自然科学基金(222300420246)
漯河医专博士后科研项目(PR20210001)。
关键词
大脑类器官
大脑皮质
诱导性多能干细胞
类器官培养
免疫荧光
小鼠
Cerebral organoid
Cerebral cortex
Induced pluripotent stem cell
Organoid culture
Immunofluorescence
Mouse
