title: Heightened potency of human pluripotent stem cell lines created by transient BMP4 exposure.
abstract: Human pluripotent stem cells (PSCs) show epiblast-type pluripotency that is maintained with ACTIVIN/FGF2 signaling. Here, we report the acquisition of a unique stem cell phenotype by both human ES cells (hESCs) and induced pluripotent stem cells (iPSCs) in response to transient (24-36 h) exposure to bone morphogenetic protein 4 (BMP4) plus inhibitors of ACTIVIN signaling (A83-01) and FGF2 (PD173074), followed by trypsin dissociation and recovery of colonies capable of growing on a gelatin substratum in standard medium for human PSCs at low but not high FGF2 concentrations. The self-renewing cell lines stain weakly for CDX2 and strongly for NANOG, can be propagated clonally on either Matrigel or gelatin, and are morphologically distinct from human PSC progenitors on either substratum but still meet standard in vitro criteria for pluripotency. They form well-differentiated teratomas in immune-compromised mice that secrete human chorionic gonadotropin (hCG) into the host mouse and include small areas of trophoblast-like cells. The cells have a distinct transcriptome profile from the human PSCs from which they were derived (including higher expression of NANOG, LEFTY1, and LEFTY2). In nonconditioned medium lacking FGF2, the colonies spontaneously differentiated along multiple lineages, including trophoblast. They responded to PD173074 in the absence of both FGF2 and BMP4 by conversion to trophoblast, and especially syncytiotrophoblast, whereas an A83-01/PD173074 combination favored increased expression of HLA-G, a marker of extravillous trophoblast. Together, these data suggest that the cell lines exhibit totipotent potential and that BMP4 can prime human PSCs to a self-renewing alternative state permissive for trophoblast development. The results may have implications for regulation of lineage decisions in the early embryo.
authors: Division of Animal Sciences, Bond Life Sciences Center and.
authors: Ying
authors: Y
authors: Yang
authors: Department of Obstetrics, Gynecology, and Women's Health, University of Missouri, Columbia, MO 65212.
authors: Katsuyuki
authors: K
authors: Adachi
authors: Department of Biochemistry, University of Missouri, Columbia, MO 65211; and.
authors: Megan A
authors: MA
authors: Sheridan
authors: Division of Animal Sciences, Bond Life Sciences Center and.
authors: Andrei P
authors: AP
authors: Alexenko
authors: Department of Obstetrics, Gynecology, and Women's Health, University of Missouri, Columbia, MO 65212.
authors: Danny J
authors: DJ
authors: Schust
authors: Department of Obstetrics, Gynecology, and Women's Health, University of Missouri, Columbia, MO 65212.
authors: Laura C
authors: LC
authors: Schulz
authors: Division of Animal Sciences, Bond Life Sciences Center and.
authors: Toshihiko
authors: T
authors: Ezashi
authors: Division of Animal Sciences, Bond Life Sciences Center and Department of Biochemistry, University of Missouri, Columbia, MO 65211; and robertsrm@missouri.edu.
authors: R Michael
authors: RM
authors: Roberts
mesh_terms: D000818:Animals; D055415:Bone Morphogenetic Protein 4 / Q000494:pharmacology*; D002454:Cell Differentiation; D002460:Cell Line; D002478:Cells, Cultured; D003094:Collagen / Q000737:chemistry; D003470:Culture Media / Q000737:chemistry; D017077:Culture Media, Conditioned; D004338:Drug Combinations; D053595:Embryonic Stem Cells / Q000378:metabolism; D005260:Female; D018507:Gene Expression Regulation, Developmental; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000378:metabolism; D007797:Laminin / Q000737:chemistry; D051379:Mice; D020411:Oligonucleotide Array Sequence Analysis; D010641:Phenotype; D010920:Placenta / Q000378:metabolism; D039904:Pluripotent Stem Cells / Q000166:cytology*; D011247:Pregnancy; D011509:Proteoglycans / Q000737:chemistry; D015398:Signal Transduction; D013724:Teratoma; D059467:Transcriptome; D014327:Trophoblasts / Q000378:metabolism
chemical_list: C525809:BMP4 protein, human; D055415:Bone Morphogenetic Protein 4; D003470:Culture Media; D017077:Culture Media, Conditioned; D004338:Drug Combinations; D007797:Laminin; D011509:Proteoglycans; C059485:matrigel; D003094:Collagen
keywords: biological sciences; developmental biology; pluripotent stem cells; totipotent; trophoblast