title: L3MBTL1 deficiency directs the differentiation of human embryonic stem cells toward trophectoderm.
abstract: Human embryonic stem cells (hESCs) can be used to study the early events in human development and, hopefully, to understand how to differentiate human pluripotent cells for clinical use. To define how L3MBTL1, a chromatin-associated polycomb group protein with transcriptional repressive activities, regulates early events in embryonic cell differentiation, we created hESC lines that constitutively express shRNAs directed against L3MBTL1. The L3MBTL1 knockdown (KD) hESCs maintained normal morphology, proliferation, cell cycle kinetics, cell surface markers, and karyotype after 40 passages. However, under conditions that promote spontaneous differentiation, the L3MBTL1 KD cells differentiated into a relatively homogeneous population of large, flat trophoblast-like cells, unlike the multilineage differentiation seen with the control cells. The differentiated L3MBTL1 KD cells expressed numerous trophoblast markers and secreted placental hormones. Although the L3MBTL1 KD cells could be induced to differentiate into various embryonic lineages, they adopted an exclusive trophoblast fate during spontaneous differentiation. Our data demonstrate that depletion of L3MBTL1 does not affect hESC self-renewal, rather it enhances differentiation toward extra-embryonic trophoblast tissues.
authors.affiliation: Molecular Pharmacology and Chemistry Program, Sloan-Kettering Institute, Memorial Sloan-Kettering Cancer Center, New York, New York 10065, USA.
authors.forename: Ruben
authors.initials: R
authors.lastname: Hoya-Arias
authors.forename: Mark
authors.initials: M
authors.lastname: Tomishima
authors.forename: Fabiana
authors.initials: F
authors.lastname: Perna
authors.forename: Francesca
authors.initials: F
authors.lastname: Voza
authors.forename: Stephen D
authors.initials: SD
authors.lastname: Nimer
mesh_terms: D000943:Antigens, Differentiation / Q000235:genetics / Q000378:metabolism; D055415:Bone Morphogenetic Protein 4 / Q000494:pharmacology / Q000502:physiology; D002454:Cell Differentiation / Q000187:drug effects; D002460:Cell Line; D049109:Cell Proliferation; D048430:Cell Shape; D002868:Chromosomal Proteins, Non-Histone / Q000172:deficiency* / Q000235:genetics; D018920:Coculture Techniques; D004475:Ectoderm / Q000166:cytology*; D053595:Embryonic Stem Cells / Q000502:physiology*; D015870:Gene Expression; D055785:Gene Knockdown Techniques; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006801:Humans; D008856:Microscopy, Fluorescence; D034622:RNA Interference; D011994:Recombinant Proteins / Q000378:metabolism; D012097:Repressor Proteins; D014327:Trophoblasts / Q000166:cytology*; D025521:Tumor Suppressor Proteins
chemical_list: D000943:Antigens, Differentiation; C525809:BMP4 protein, human; D055415:Bone Morphogenetic Protein 4; D002868:Chromosomal Proteins, Non-Histone; C120887:L3MBTL1 protein, human; D011994:Recombinant Proteins; D012097:Repressor Proteins; D025521:Tumor Suppressor Proteins; D049452:Green Fluorescent Proteins