The molecular mechanisms controlling mouse embryonic stem cell(ESC)metastability,i.e.their capacity to fluctuate between different states of pluripotency,are not fully resolved.We developed and used a novel automation...The molecular mechanisms controlling mouse embryonic stem cell(ESC)metastability,i.e.their capacity to fluctuate between different states of pluripotency,are not fully resolved.We developed and used a novel automation platform,the Cellmaker,to screen a library of metabolites on two ESC-based phenotypic assays(i.e.proliferation and colony phenotype)and identified two metabolically related amino acids,namely L-proline(L-Pro)and L-ornithine(L-Orn),as key regulators of ESC metastability.Both compounds,but mainly L-Pro,force ESCs toward a novel epiblast stem cell(EpiSC)-like state,in a dose-and time-dependent manner.Unlike EpiSCs,L-Pro-induced cells(PiCs)contribute to chimeric embryos and rely on leukemia inhibitor factor(LIF)to self-renew.Furthermore,PiCs revert to ESCs or differentiate randomly upon removal of either L-Pro or LIF,respectively.Remarkably,PiC generation depends on both L-Pro metabolism(uptake and oxidation)and Fgf5 induction,and is strongly counteracted by antioxidants,mainly L-ascorbic acid(vitamin C,Vc).ESCs↔PiCs phenotypic transition thus represents a previously undefined dynamic equilibrium between pluripotent states,which can be unbalanced either toward an EpiSC-like or an ESC phenotype by L-Pro/L-Orn or Vc treatments,respectively.All together,our data provide evidence that ESC metastability can be regulated at a metabolic level.展开更多
基金This work was supported by Associazione Italiana Ricerca sul Cancro[IG-6128 to G.M.and IG-4840 to S.D.F.]Telethon[GGP-08120 to G.M.]+1 种基金Regione Campania-Programma Operativo Regionale[CRdC WP11 to S.F.,S.D.F,D.D.C.,G.M.and E.J.P.]Ministero Istruzione Universita Ricerca[Medical Research in Italy RBNE08HM7T_003 to E.J.P.].
文摘The molecular mechanisms controlling mouse embryonic stem cell(ESC)metastability,i.e.their capacity to fluctuate between different states of pluripotency,are not fully resolved.We developed and used a novel automation platform,the Cellmaker,to screen a library of metabolites on two ESC-based phenotypic assays(i.e.proliferation and colony phenotype)and identified two metabolically related amino acids,namely L-proline(L-Pro)and L-ornithine(L-Orn),as key regulators of ESC metastability.Both compounds,but mainly L-Pro,force ESCs toward a novel epiblast stem cell(EpiSC)-like state,in a dose-and time-dependent manner.Unlike EpiSCs,L-Pro-induced cells(PiCs)contribute to chimeric embryos and rely on leukemia inhibitor factor(LIF)to self-renew.Furthermore,PiCs revert to ESCs or differentiate randomly upon removal of either L-Pro or LIF,respectively.Remarkably,PiC generation depends on both L-Pro metabolism(uptake and oxidation)and Fgf5 induction,and is strongly counteracted by antioxidants,mainly L-ascorbic acid(vitamin C,Vc).ESCs↔PiCs phenotypic transition thus represents a previously undefined dynamic equilibrium between pluripotent states,which can be unbalanced either toward an EpiSC-like or an ESC phenotype by L-Pro/L-Orn or Vc treatments,respectively.All together,our data provide evidence that ESC metastability can be regulated at a metabolic level.
