Multiple factors to assist human-derived induced pluripotent stem cells to efficiently differentiate into midbrain dopaminergic neurons  

作　　者：Yalan Chen Junxin Kuang Yimei Niu Hongyao Zhu Xiaoxia Chen Kwok-Fai So Anding Xu Lingling Shi 

机构地区：[1]Guangdong-Hong Kong-Macao Institute of CNS Regeneration,Ministry of Education CNS Regeneration Collaborative Joint Laboratory,Jinan University [2]Department of Neurology and Stroke Center,The First Affiliated Hospital,Jinan University [3]Department of Psychiatry,The First Affiliated Hospital,Jinan University [4]Co-innovation Center of Neuroregeneration,Nantong University

出　　处：《Neural Regeneration Research》2024年第4期908-914,共7页中国神经再生研究（英文版）

基　　金：supported by the National Natural Science Foundation of China,No.81771222(to LS);Guangzhou Key Research Program on Brain Science,Nos.202007030011,202206060001(to LS);the Program of Introducing Talents of Discipline to Universities of China,No.B14036(to KFS)。

摘　　要：Midbrain dopaminergic neurons play an important role in the etiology of neurodevelopmental and neurodegenerative diseases.They also represent a potential source of transplanted cells for therapeutic applications.In vitro differentiation of functional midbrain dopaminergic neurons provides an accessible platform to study midbrain neuronal dysfunction and can be used to examine obstacles to dopaminergic neuronal development.Emerging evidence and impressive advances in human induced pluripotent stem cells,with tuned neural induction and differentiation protocols,makes the production of induced pluripotent stem cell-derived dopaminergic neurons feasible.Using SB431542 and dorsomorphin dual inhibitor in an induced pluripotent stem cell-derived neural induction protocol,we obtained multiple subtypes of neurons,including 20%tyrosine hydroxylase-positive dopaminergic neurons.To obtain more dopaminergic neurons,we next added sonic hedgehog(SHH)and fibroblast growth factor 8(FGF8)on day 8 of induction.This increased the proportion of dopaminergic neurons,up to 75%tyrosine hydroxylase-positive neurons,with 15%tyrosine hydroxylase and forkhead box protein A2(FOXA2)co-expressing neurons.We further optimized the induction protocol by applying the small molecule inhibitor,CHIR99021(CHIR).This helped facilitate the generation of midbrain dopaminergic neurons,and we obtained 31-74%midbrain dopaminergic neurons based on tyrosine hydroxylase and FOXA2 staining.Thus,we have established three induction protocols for dopaminergic neurons.Based on tyrosine hydroxylase and FOXA2 immunostaining analysis,the CHIR,SHH,and FGF8 combined protocol produces a much higher proportion of midbrain dopaminergic neurons,which could be an ideal resource for tackling midbrain-related diseases.

关 键 词：dopaminergic neurons FGF signal induced pluripotent stem cells MIDBRAIN neural differentiation SHH signal SMAD signal WNT signal 

分 类 号：R741[医药卫生—神经病学与精神病学]