title: Targeting SOX17 in human embryonic stem cells creates unique strategies for isolating and analyzing developing endoderm.
abstract: Human embryonic stem cells (hESCs) can provide insights into development of inaccessible human tissues such as embryonic endoderm. Progress in this area has been hindered by a lack of methods for isolating endodermal cells and tracing fates of their differentiated progeny. By using homologous recombination in human ESCs, we inserted an enhanced green fluorescent protein (eGFP) transgene into the SOX17 locus, a postulated marker of human endoderm. FACS purification and gene expression profiling confirmed that SOX17(+)-hESC progeny expressed endodermal markers and unveiled specific cell surface protein combinations that permitted FACS-based isolation of primitive gut tube endodermal cells produced from unmodified human ESCs and from induced pluripotent stem cells (iPSC). Differentiating SOX17(+) endodermal cells expressed markers of liver, pancreas, and intestinal epithelium in vitro and gave rise to endodermal progeny in vivo. Thus, prospective isolation, lineage tracing, and developmental studies of SOX17(+) hESC progeny have revealed fundamental aspects of human endodermal biology.
authors.affiliation: Department of Developmental Biology, Stanford University School of Medicine, Stanford, CA 94305, USA.
authors.forename: Pei
authors.initials: P
authors.lastname: Wang
authors.forename: Ryan T
authors.initials: RT
authors.lastname: Rodriguez
authors.forename: Jing
authors.initials: J
authors.lastname: Wang
authors.forename: Amar
authors.initials: A
authors.lastname: Ghodasara
authors.forename: Seung K
authors.initials: SK
authors.lastname: Kim
mesh_terms: D015415:Biomarkers / Q000378:metabolism; D002454:Cell Differentiation / Q000235:genetics; D002460:Cell Line; D019070:Cell Lineage / Q000235:genetics; D002462:Cell Membrane / Q000378:metabolism; D002469:Cell Separation / Q000379:methods*; D053595:Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism*; D004707:Endoderm / Q000166:cytology* / Q000196:embryology*; D041981:Gastrointestinal Tract / Q000166:cytology / Q000196:embryology; D018507:Gene Expression Regulation, Developmental; D018390:Gene Targeting; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006801:Humans; D020411:Oligonucleotide Array Sequence Analysis; D011995:Recombination, Genetic / Q000235:genetics; D055760:SOXF Transcription Factors / Q000235:genetics* / Q000378:metabolism
chemical_list: D015415:Biomarkers; C109188:SOX17 protein, human; D055760:SOXF Transcription Factors; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins