Generation of a human haploid neural stem cell line for genome-wide genetic screening  

作　　者：Hai-Song Wang Xin-Rui Ma Wen-Bin Niu Hao Shi Yi-Dong Liu Ning-Zhao Ma Nan Zhang Zi-Wei Jiang Ying-Pu Sun 

机构地区：[1]Center for Reproductive Medicine,The First Affiliated Hospital of Zhengzhou University,Zhengzhou University,Zhengzhou 450052,Henan Province,China [2]Basic Medical School,Zhengzhou University,Zhengzhou 450052,Henan Province,China

出　　处：《World Journal of Stem Cells》2023年第7期734-750,共17页世界干细胞杂志（英文版）（电子版）

基　　金：Supported by the National Natural Science Foundation of China,No.81901476.

摘　　要：BACKGROUND Haploid embryonic stem cells(haESCs)have been established in many species.Differentiated haploid cell line types in mammals are lacking due to spontaneous diploidization during differentiation that compromises lineage-specific screens.AIM To derive human haploid neural stem cells(haNSCs)to carry out lineage-specific screens.METHODS Human haNSCs were differentiated from human extended haESCs with the help of Y27632(ROCK signaling pathway inhibitor)and a series of cytokines to reduce diploidization.Neuronal differentiation of haNSCs was performed to examine their neural differentiation potency.Global gene expression analysis was conducted to compare haNSCs with diploid NSCs and haESCs.Fluorescence activated cell sorting was performed to assess the diploidization rate of extended haESCs and haNSCs.Genetic manipulation and screening were utilized to evaluate the significance of human haNSCs as genetic screening tools.RESULTS Human haESCs in extended pluripotent culture medium showed more compact and smaller colonies,a higher efficiency in neural differentiation,a higher cell survival ratio and higher stability in haploidy maintenance.These characteristics effectively facilitated the derivation of human haNSCs.These human haNSCs can be generated by differentiation and maintain haploidy and multipotency to neurons and glia in the long term in vitro.After PiggyBac transfection,there were multiple insertion sites in the human haNSCs’genome,and the insertion sites were evenly spread across all chromosomes.In addition,after the cells were treated with manganese,we were able to generate a list of manganese-induced toxicity genes,demonstrating their utility as genetic screening tools.CONCLUSION This is the first report of a generated human haploid somatic cell line with a complete genome,proliferative ability and neural differentiation potential that provides cell resources for recessive inheritance and drug targeted screening.

关 键 词：HAPLOID Neural stem cells Extended pluripotency Genetic screening Cell therapy 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]