biostudies-literatureSoncin FEunice Kennedy Shriver National Institute of Child Health and Human DevelopmentNICHD NIH HHSNCATS NIH HHSUniversity of California, San DiegoNational Institutes of HealthCalifornia Institute for Regenerative MedicineAmerican Society for Reproductive MedicineNIH HHSNIGMS NIH HHSNational Institute of Child Health and Human DevelopmentNational Science Foundation1303-1317https://www.ebi.ac.uk/biostudies/studies/S-EPMC9214048biostudies-literatureUnknown17(6)Trophoblast stem cells (TSCs) have recently been derived from human embryos and early-first-trimester placenta; however, aside from ethical challenges, the unknown disease potential of these cells limits their scientific utility. We have previously established a bone morphogetic protein 4 (BMP4)-based two-step protocol for differentiation of primed human pluripotent stem cells (hPSCs) into functional trophoblasts; however, those trophoblasts could not be maintained in a self-renewing TSC-like state. Here, we use the first step from this protocol, followed by a switch to newly developed TSC medium, to derive bona fide TSCs. We show that these cells resemble placenta- and naive hPSC-derived TSCs, based on their transcriptome as well as their in vitro and in vivo differentiation potential. WeStem cell reportsDerivation of functional trophoblast stem cells from primed human pluripotent stem cells.PMC9214048TG-MCB140074T32 GM008806K99 HD091452UL1 TR0014422UL1TR001442-06R01 HD089537R00 HD091452EDUC2-08388R01 HD096260R01HD-089537R01-HD096260T32 GM007198S10 OD026929K99-R00-HD091452T32GM8806ACI-1548562Cheung VCKallol SFarah OKittle RJackson MGMorey RFisch KMSoncin FBui TPizzo DParast MMRequena DFMeads MHorii MChousal JfalseDerivation of functional trophoblast stem cells from primed human pluripotent stem cells.Trophoblast stem cells (TSCs) have recently been derived from human embryos and early-first-trimester placenta; however, aside from ethical challenges, the unknown disease potential of these cells limits their scientific utility. We have previously established a bone morphogetic protein 4 (BMP4)-based two-step protocol for differentiation of primed human pluripotent stem cells (hPSCs) into functional trophoblasts; however, those trophoblasts could not be maintained in a self-renewing TSC-like state. Here, we use the first step from this protocol, followed by a switch to newly developed TSC medium, to derive bona fide TSCs. We show that these cells resemble placenta- and naive hPSC-derived TSCs, based on their transcriptome as well as their in vitro and in vivo differentiation potential. We2022-01-01T00:00:00Z2022 Jun2025-04-04T03:06:18.188Z2025-02-19T05:06:17.931ZS-EPMC92140483559485810.1016/j.stemcr.2022.04.013