Canonical and early lineage-specific stem cell types identified in planarian SirNeoblasts  

作　　者：Kaimeng Niu Hao Xu Yuanyi Zhou Xiong Yun Zhao Chong Gao Chris WSeidel Xue Pan Yuqing Ying Kai Lei 

机构地区：[1]Westlake Laboratory of Life Sciences and Biomedicine,Key Laboratory of Growth Regulation and Translational Research of Zhejiang Province,School of Life Sciences,Westlake University,Hangzhou,Zhejiang,China. [2]Institute of Biology,Westlake Institute for Advanced Study,Hangzhou,Zhejiang,China [3]College of Life Sciences,Zhejiang University,Hangzhou,Zhejiang,China [4]Stowers Institute for Medical Research,Kansas City,MO 64110,USA

出　　处：《Cell Regeneration》2021年第1期177-187,共11页细胞再生（英文）

基　　金：K.L.is supported by the National Natural Science Foundation of China(31970750);the start-up fund from the Westlake Education Foundation.

摘　　要：Background:The pluripotent stem cells in planarians,a model for tissue and cellular regeneration,remain further identification.We recently developed a method to enrich piwi-1+cells in Schmidtea mediterranea,by staining cells with SiR-DNA and Cell Tracker Green,named SirNeoblasts that permits their propagation and subsequent functional study in vivo.Since traditional enrichment for planarian neoblasts by Hoechst 33342 staining generates X1 cells,blocking the cell cycle and inducing cytotoxicity,this method by SiR-DNA and Cell Tracker Green represents a complementary technological advance for functional investigation of cell fate and regeneration.However,the similarities in heterogeneity of cell subtypes between SirNeoblasts and X1 remain unknown.Results:In this work,we performed single cell RNA sequencing of SirNeoblasts for comparison with differential expression patterns in a publicly available X1 single cell RNA sequencing data.We found first that all of the lineagespecific progenitor cells in X1 were present in comparable proportions in SirNeoblasts.In addition,SirNeoblasts contain an early muscle progenitor that is unreported in X1.Analysis of new markers for putative pluripotent stem cells identified here,with subsequent sub-clustering analysis,revealed earlier lineages of epidermal,muscular,intestinal,and pharyngeal progenitors than have been observed in X1.Using the gcm as a marker,we also identified a cell subpopulation resided in previously identified tgs-1+neoblasts.Knockdown of gcm impaired the neoblast repopulation,suggesting a function of gcm in neoblasts.Conclusions:In summary,the use of SirNeoblasts will enable broad experimental advances in regeneration and cell fate specification,given the possibility for propagation and transplantation of recombinant and mutagenized pluripotent stem cells that are not previously afforded to this rapid and versatile model system.

关 键 词：PLANARIA Regeneration NEOBLAST SiR-DNA scRNA-seq 

分 类 号：R73[医药卫生—肿瘤]