Human naive pluripotent stem cells(PSCs)hold great promise for embryonic development studies.Existing induction and culture strategies for these cells,heavily dependent on MEK inhibitors,lead to widespread DNA hypomet...Human naive pluripotent stem cells(PSCs)hold great promise for embryonic development studies.Existing induction and culture strategies for these cells,heavily dependent on MEK inhibitors,lead to widespread DNA hypomethylation,aberrant imprinting loss,and genomic instability during extended culture.Here,employing high-content analysis alongside a bifluorescence reporter system indicative of human naive pluripotency,we screened over 1,600 chemicals and identified seven promising candidates.From these,we developed four optimized media-LAY,LADY,LUDY,and LKPY-that effectively induce and sustain PSCs in the naive state.Notably,cells reset or cultured in these media,especially in the LAY system,demonstrate improved genome-wide DNA methylation status closely resembling that of pre-implantation counterparts,with partially restored imprinting and significantly enhanced genomic stability.Overall,our study contributes advancements to naive pluripotency induction and long-term maintenance,providing insights for further applications of naive PSCs.展开更多
基金The bulk RNA-seq datasets generated in this study are available at GEO:GSE252312The WGBS datasets generated in this study are available at GEO:GSE252313+2 种基金The accession numbers for the RNA-seq data of human embryos are E-MTAB-3929 and GSE36552The accession numbers for the RNA-seq data of published cell lines areE-MTAB-2856,GSE59435,GSE166401,CNP0001454,GSE150772,and GSE174771The accession numbers for the WGBS data are GSE49828,JGAS00000000006,GSE60945,GSE76970,GSE90168,CNP0001454,and GSE142812。
文摘Human naive pluripotent stem cells(PSCs)hold great promise for embryonic development studies.Existing induction and culture strategies for these cells,heavily dependent on MEK inhibitors,lead to widespread DNA hypomethylation,aberrant imprinting loss,and genomic instability during extended culture.Here,employing high-content analysis alongside a bifluorescence reporter system indicative of human naive pluripotency,we screened over 1,600 chemicals and identified seven promising candidates.From these,we developed four optimized media-LAY,LADY,LUDY,and LKPY-that effectively induce and sustain PSCs in the naive state.Notably,cells reset or cultured in these media,especially in the LAY system,demonstrate improved genome-wide DNA methylation status closely resembling that of pre-implantation counterparts,with partially restored imprinting and significantly enhanced genomic stability.Overall,our study contributes advancements to naive pluripotency induction and long-term maintenance,providing insights for further applications of naive PSCs.
