Human naïve 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 h...Human naïve 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 naïve pluripotency,we screened over 1,600chemicals 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 naïve 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 naïve pluripotency induction and long-term maintenance,providing insights for further applications of naïve PSCs.展开更多
基金supported by the Ministry of Science and Technology of China (Grant No. 2021YFA11020000 to Y.W., Grant No. 2022YFC2702200 to S.G. and Grant No. 2023YFA1800019 to Y.W.)the National Key R&D Program of China (Grant No. 2024YFA1107000 to Y.B.)+5 种基金the National Natural Science Foundation of China (Grant Nos. 32270850 and 32470843 to Y.W., Grant Nos. 32488101 and 92168205 to S.G., Grant Nos. 32370842 and 32100633 to Y.B.)the Shuguang Program of Shanghai Education Development Foundation and Shanghai Municipal Education Commission (Grant No. 22SG20 to Y.W.)the Chenguang Program of Shanghai Education Development Foundation and Shanghai Municipal Education Commission (Grant No. 24CGA22 to Y.B.)the Science and Technology Commission of Shanghai Municipality (Grant No. 21JC1405500 to S.G.)the Fundamental Research Funds for the Central Universities (Grant No. 22120240319 to Y.B., Grant No. 22120240255 to Y.W. and Grant No. 22120250374)the Peak Disciplines (Type IV) of Institutions of Higher Learning in Shanghai.
文摘Human naïve 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 naïve pluripotency,we screened over 1,600chemicals 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 naïve 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 naïve pluripotency induction and long-term maintenance,providing insights for further applications of naïve PSCs.
