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

doi:10.16098/j.issn.0529-1356.2017.04.004

Abstract

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

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.

References

［1］Mizumoto H, Ishihara K, Nakazawa K, et al. A new culture technique for hepatocyte organoid formation and longterm maintenance of liver-specific functions［J］. Tissue Eng Part C Methods,2008, 14(2):167-175.

［2］Pastula A, Middelhoff M, Brandtner A, et al. Three-dimensional gastrointestinal organoid culture in combination with nerves or fibroblasts: a method to characterize the gastrointestinal stem cell niche［J］. Stem Cells Int, 2016, 2016:3710836.

［3］Takahashi K, Tanabe K, Ohnuki M, et al. Induction of pluripotent stem cells from adult human fibroblasts by defined factors［J］. Cell, 2007, 131(5):861-872.

［4］Takahashi K, Yamanaka S. Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors［J］. Cell,2006, 126(4):663-676.

［5］Eiraku M, Watanabe K, Matsuo-Takasaki M, et al. Self-organized formation of polarized cortical tissues from ESCs and its active manipulation by extrinsic signals［J］. Cell Stem Cell, 2008, 3(5):519-532.

［6］Nakano T, Ando S, Takata N, et al. Self-formation of optic cups and storable stratified neural retina from human ESCs［J］. Cell Stem Cell, 2012, 10(6):771-785.

［7］Elkabetz Y, Panagiotakos G, Al Shamy G, et al. Human ES cell-derived neural rosettes reveal a functionally distinct early neural stem cell stage［J］. Genes Dev, 2008, 22(2):152-165.

［8］Zhang Y, Niu B, Yu D, et al. Radial glial cells and the lamination of the cerebellar cortex［J］. Brain Struct Funct,2010, 215(2):115-122.

［9］Howard BM, Zhicheng M, Filipovic R, et al. Radial glia cells in the developing human brain［J］. Neuroscientist,2008, 14(5):459-473.

［10］Than-Trong E, Bally-Cuif L. Radial glia and neural progenitors in the adult zebrafish central nervous system［J］. Glia, 2015, 63(8):1406-1428.

［11］Sild M, Ruthazer ES. Radial glia: progenitor, pathway, and partner［J］. Neuroscientist, 2011, 17(3):288-302.

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

［13］Chen B, Wang SS, Hattox AM, et al. The Fezf2-Ctip2 genetic pathway regulates the fate choice of subcortical projection neurons in the developing cerebral cortex［J］. Proc Natl Acad Sci USA, 2008, 105(32):11382-11387.

［14］Gomez-Lopez S, Wiskow O, Favaro R, et al. Sox2 and Pax6 maintain the proliferative and developmental potential of gliogenic neural stem cells In vitro［J］. Glia, 2011, 59(11):1588-1599.

［15］Frotscher M. Cajal-Retzius cells, Reelin, and the formation of layers［J］. Curr Opin Neurobiol,1998, 8(5):570-575.

［16］Frotscher M. Dual role of Cajal-Retzius cells and reelin in cortical development［J］. Cell Tissue Res, 1997, 290(2):315-322.

［17］Wu P, Li MS, Yu DM, et al. Reelin, a guidance signal for the regeneration of the entorhino-hippocampal path［J］. Brain Res, 2008, 1208:1-7.

［18］Fan WJ，Chen XD，Yuan L，et al. Neural differentiation and synapse formation in mouse induced pluripotent stem cells［J］. Acta Anatamica Sinica,2015,46(1):6-12.(in Chinese)范文娟，陈旭东，袁磊，等. 小鼠诱导性多能干细胞的神经细胞分化与突触连接的建立及其功能分析［J］. 解剖学报, 2015, 46(1): 6-12.

［19］Fu S, Shi ZhY, Fan WJ, et al. Microenvironments induce iPSCs and BMSCs into neuron-like cells——Reelin’s regulative role in cell differentiation and polarization［J］. Acta Physiologica Sinica, 2015, 67(4): 357-369.(in Chinese)

付苏，石贞玉，范文娟，等. 微环境诱导iPSCs和BMSCs向神经元样细胞分化——Reelin对细胞分化和极性化的调节作用［J］.生理学报, 2015, 67(4): 357-369.

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