体外培养大脑皮质类器官与活体大脑皮质发育的比较

doi:10.16098/j.issn.0529-1356.2023.04.002

解剖学报 ›› 2023, Vol. 54 ›› Issue (4): 383-391.doi: 10.16098/j.issn.0529-1356.2023.04.002

体外培养大脑皮质类器官与活体大脑皮质发育的比较

范文娟^1,2 陈旭东¹ 陈永芳¹杨旭光¹ 金少举¹ 赵志军^1* 邓锦波^1*

1.漯河医学高等专科学校，河南省营养与健康工程研究中心，河南漯河 462002;2.漯河市第一人民医院神经内科，河南漯河 462000

收稿日期:2022-02-17 修回日期:2022-04-17 出版日期:2023-08-06 发布日期:2023-08-06
通讯作者: 赵志军; 邓锦波 E-mail:zzj20220216@163.com
基金资助:
河南省科技厅项目;河南省科技厅项目;河南省自然科学基金

Developmental comparison between cerebral organoids in vitro and body’s cortices in vivo

FAN Wen-juan^1,2 CHEN Xu-dong¹ CHEN Yong-fang¹ YANG Xu-guang¹ JIN Shao-ju¹ZHAO Zhi-jun^1* DENG Jin-bo^1*

1. Luohe Medical College, He’nan Province Engineering Research Center of Nutrition and Health, He’nan Luohe 462002, China ;
2. Neurology Department, the First People’s Hospital of Luohe City, He’nan Luohe 462000, China

Received:2022-02-17 Revised:2022-04-17 Online:2023-08-06 Published:2023-08-06
Contact: ZHAO Zhi-jun; DENG Jin-bo E-mail:zzj20220216@163.com

摘要/Abstract

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。

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

Abstract:

Objective To understand the characteristics and developmental differences between cerebral organoids in vitro and normal cerebral cortices in vivo. Methods 1. Grouping: cerebral cortices in vivo group and cultured cerebral organoids in vitro group. 2. Sample collection: cortical tissues were collected from Kunming mouse embryos at embryonic day 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 different culture time point were collected, and more than 3 samples were collected at each time point. 3. Detection method: the distribution of different types of cells in each group of specimens was analyzed by immunofluorescent staining. Results While relative similarities between in vivo cerebral cortical development and the cerebral organoids in vitro were observed, including the histogenesis, and the morphological differentiation of nerve cells and glial cells, the lamellar architecture of cerebral cortex in mouse brain was not observed in cerebral organoids. Conclusion The development of cerebral organoids in vitro has some similarity with body’s cortical development. Therefore, cerebral organoids can be used to a substitution of cortex and diseases’ models, but improvement of the existing technologies is necessary.

Key words: Cerebral organoid, Cerebral cortex, Induced pluripotent stem cell, Organoid culture, Immunofluorescence, Mouse

中图分类号:

R329

范文娟陈旭东陈永芳杨旭光金少举赵志军邓锦波. 体外培养大脑皮质类器官与活体大脑皮质发育的比较[J]. 解剖学报, 2023, 54(4): 383-391.

FAN Wen-juan CHEN Xu-dong CHEN Yong-fang YANG Xu-guang JIN Shao-ju ZHAO Zhi-jun DENG Jin-bo. Developmental comparison between cerebral organoids in vitro and body’s cortices in vivo[J]. Acta Anatomica Sinica, 2023, 54(4): 383-391.

参考文献

［1］ Shi Y, Wu Q, Wang X. Modeling brain development and diseases with human cerebral organoids［J］. Curr Opin Neurobiol, 2021, 66: 103-115.

［2］Lancaster MA, Renner M, Martin CA, et al. Cerebral organoids model human brain development and microcephaly［J］. Nature, 2013, 501(7467): 373-379.

［3］Velasco S, Kedaigle AJ, Simmons SK, et al. Individual brain organoids reproducibly form cell diversity of the human cerebral cortex［J］. Nature, 2019, 570(7762): 523-527.

［4］Li Y, Muffat J, Omer A, et al. Induction of Expansion and Folding in Human Cerebral Organoids［J］. Cell Stem Cell, 2017, 20(3): 385-396.

［5］Fan WJ，Wang Q，Sun YZh, et al. Mouse induced pluripotent stem cell-derived cortical organoids and its biological characteristics［J］. Acta Anatomica Sinica,2017,48(4):387-396. (in Chinese)

范文娟,王倩,孙仪征,等.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.

［6］Xiong Y, Chen PH, Zhou Y. Neurobiology ［M］. 2nd ed. Beijing: Science Press, 2021:108-118. (in Chinese)

熊鹰，陈鹏慧，周艺. 神经生物学［M］. 第2版. 北京：科学出版社，2021:108-118.

［7］Takamori Y, Mori T, Wakabayashi T, et al. Nestin-positive microglia in adult rat cerebral cortex［J］. Brain Res, 2009, 1270: 10-18.

［8］Chew L, Aonuevo A, Knock E. Generating cerebral organoids from human pluripotent stem cells［J］. Methods Mol Biol, 2022, 2389: 177-199.

［9］Lancaster MA, Corsini NS, Wolfinger S, et al. Guided self-organization and cortical plate formation in human brain organoids［J］. Nat Biotechnol, 2017, 35(7): 659-666.

［10］Sidney LE, Branch MJ, Hopkinson A, et al. Concise review: evidence for CD34 as a common marker for diverse progenitors［J］. Stem Cells, 2014, 32(6):1380-1389.

［11］Qian X, Su Y, Adam CD, et al. Sliced human cortical organoids for modeling distinct cortical layer formation［J］. Cell Stem Cell, 2020, 26(5): 766-781.

［12］Qian X, Nguyen HN, Song MM, et al. Brain-region-specific organoids using mini-bioreactors for modeling ZIKV exposure［J］. Cell, 2016, 165(5): 1238-1254.

［13］Renner M, Lancaster MA, Bian S, et al. Self-organized developmental patterning and differentiation in cerebral organoids［J］. EMBO J, 2017, 36(10): 1316-1329.

［14］Fan W, Sun Y, Shi Z, et al. Mouse induced pluripotent stem cells-derived Alzheimer’s disease cerebral organoid culture and neural differentiation disorders［J］. Neurosci Lett, 2019, 711: 134433.

［15］Yakoub AM. Cerebral organoids exhibit mature neurons and astrocytes and recapitulate electrophysiological activity of the human brain［J］. Neural Regen Res, 2019, 14(5): 757-761.

［16］Giandomenico SL, Mierau SB, Gibbons GM, et al. Cerebral organoids at the air-liquid interface generate diverse nerve tracts with functional output［J］. Nat Neurosci, 2019, 22(4): 669-679.

［17］Rash BG, Grove EA. Area and layer patterning in the developing cerebral cortex［J］. Curr Opin Neurobiol, 2006, 16(1): 25-34.

［18］Ha S, Tripathi PP, Daza RA, et al. Reelin mediates hippocampal Cajal-Retzius cell positioning and infrapyramidal blade morphogenesis［J］. J Dev Biol, 2020, 8(3):20.

［19］Causeret F, Moreau MX, Pierani A, et al. The multiple facets of Cajal-Retzius neurons［J］. Development, 2021, 148(11):199409.

［20］Fan WJ, Cheng WJ, Niu YL, et al. Normal development of Cajal-Retzius cells in mouse hippocampus and their changes in APPswe transgenic mice［J］. Acta Anatomica Sinica, 2010, 41(2): 211-218. (in Chinese)

范文娟, 程维杰, 牛艳丽等. 海马Cajal-Retzius细胞的正常发育及在APPswe转基因小鼠中的改变［J］.解剖学报, 2010, 41(2): 211-218.

［21］Gabriel E, Albanna W, Pasquini G, et al. Human brain organoids assemble functionally integrated bilateral optic vesicles［J］. Cell Stem Cell, 2021, 28(10): 1740-1757.

［22］Cakir B, Xiang Y, Tanaka Y, et al. Engineering of human brain organoids with a functional vascular-like system［J］. Nat Methods, 2019,16(11):1169-1175.

［23］Ahn Y, An JH, Yang HJ, et al. Human blood vessel organoids penetrate human cerebral organoids and form a vessel-like system［J］. Cells, 2021,10(8):2036.

[1]	洪晓敏李三强崔钦奕郑润月杨孟利罗仁利李前辉 . 酒精性肝纤维化核因子κB信号通路与性别的差异 [J]. 解剖学报, 2024, 55(1): 55-61.
[2]	袁蕾杨智伟杨惠刘政海何洁万炜. 三七总皂苷抑制小鼠星形胶质细胞活化缓解完全弗氏佐剂所致炎性痛 [J]. 解剖学报, 2024, 55(1): 25-31.
[3]	马婷婷陈建红刘爱翠李海宁. 抑制lncRNA TUG1下调核苷酸结合寡聚结构域样受体蛋白1炎症小体在延缓阿尔茨海默病进展的作用 [J]. 解剖学报, 2024, 55(1): 32-42.
[4]	邹成功冯浩陈兵唐辉邵川孙谋杨荣何家全. 创伤性颅脑损伤中神经功能障碍与认知功能损伤的动态变化 [J]. 解剖学报, 2024, 55(1): 43-48.
[5]	苏芃王兴仪梁景岩熊天庆. 一种低密度及高纯度胎鼠原代海马神经元培养方法的优化及鉴定[J]. 解剖学报, 2024, 55(1): 113-119.
[6]	许亚平余悦欣陈金福王柯柯郭志坤 . 高龄大鼠心室肌间质弹性纤维和胶原纤维的结构分布特征及其机制 [J]. 解剖学报, 2023, 54(6): 716-721.
[7]	刘丽平朱梦妮徐慧 . 白细胞介素6对脂多糖诱导所致类子痫前期大鼠有核红细胞的影响 [J]. 解剖学报, 2023, 54(6): 722-729.
[8]	张琴杨俊霞蒋青松. 福辛普利对糖尿病视网膜病变小鼠血管紧张素转化酶2和血管内皮生长因子的影响[J]. 解剖学报, 2023, 54(6): 628-634.
[9]	陈子璇司丽娜舒薇菡张欣魏晨阳程露阳杨松鹤乔跃兵. 褪黑素调控下丘脑PI3K/Akt/mTOR信号延缓雌性小鼠青春期启动的机制 [J]. 解剖学报, 2023, 54(6): 644-651.
[10]	刘哲川李坤马帅男孟家琪王艳芹. 利拉鲁肽对百草枯诱导的小鼠帕金森病模型炎症及线粒体融合/分裂的影响 [J]. 解剖学报, 2023, 54(6): 676-681.
[11]	刘叶楚亚楠徐岑璐何佳澄苏炳银太颢然. Roscovitine挽救帕金森病小鼠相关脑区神经元丢失和神经炎症[J]. 解剖学报, 2023, 54(6): 635-643.
[12]	魏纯纯王平林方兴麻献华章卫平谢志芳 . 小鼠棕色脂肪透射电子显微镜样品固定方法的改良 [J]. 解剖学报, 2023, 54(6): 738-743.
[13]	阎锟武筱林刘英娟葛科立任雷鸣李红云 . 脑蛋白水解物-Ⅰ对帕金森病小鼠肠道菌群的调节作用[J]. 解剖学报, 2023, 54(5): 497-504.
[14]	李梦一吴安婷许泽婷张庭李军伟周鹏崔怀瑞孙臣友 . 哺乳动物雷帕霉素靶蛋白复合物2/Akt信号通路对6-羟基多巴胺处理的SH-SY5Y细胞模型的作用及分子机制[J]. 解剖学报, 2023, 54(5): 521-530.
[15]	顾玲张萍宋科昕李娜陈虹宇张婕丛敬翟效月 . 发生发育小鼠肾输尿管芽分支的形态特点[J]. 解剖学报, 2023, 54(5): 593-598.

体外培养大脑皮质类器官与活体大脑皮质发育的比较

Developmental comparison between cerebral organoids in vitro and body’s cortices in vivo

PDF

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价