title: Transcriptional profiling of human embryonic stem cells differentiating to definitive and primitive endoderm and further toward the hepatic lineage.
abstract: Human embryonic stem cells (hESC) can differentiate into a variety of specialized cell types, and they constitute a useful model system to study embryonic development in vitro. In order to fully utilize the potential of these cells, the mechanisms that regulate the developmental processes of specific lineage differentiation need to be better defined. The aim of this study was to explore the molecular program involved in the differentiation of hESC toward definitive endoderm (DE) and further into the hepatic lineage, and to compare that with primitive endoderm (PrE) differentiation. To that end, we applied two protocols: a specific DE differentiation protocol and an intrinsic differentiation protocol that mainly mediates PrE formation. We collected hESC, hESC-derived DE, DE-derived hepatocyte-progenitors (DE-Prog), DE-derived hepatocyte-like cells (DE-Hep), and the corresponding PrE derivatives. The samples were analyzed using microarrays, and we identified sets of genes that were exclusively up-regulated in DE derivatives (compared to PrE derivatives) at discrete developmental stages. We also investigated known protein interactions among the set of up-regulated genes in DE-Hep. The results demonstrate important differences between DE and PrE differentiation on the transcriptional level. In particular, our results identify a unique molecular program, exclusively activated during development of DE and the subsequent differentiation of DE toward the hepatic lineage. We identified key genes and pathways of potential importance for future efforts to improve hepatic differentiation from hESC. These results reveal new opportunities for rational design of specific interventions with the purpose of generating enriched populations of DE derivatives, including functional hepatocytes.
authors.affiliation: School of Life Sciences, University of Skövde , Skövde, Sweden. jane.synnergren@his.se
authors.forename: Jane
authors.initials: J
authors.lastname: Synnergren
authors.forename: Nico
authors.initials: N
authors.lastname: Heins
authors.forename: Gabriella
authors.initials: G
authors.lastname: Brolén
authors.forename: Gustav
authors.initials: G
authors.lastname: Eriksson
authors.forename: Anders
authors.initials: A
authors.lastname: Lindahl
authors.forename: Johan
authors.initials: J
authors.lastname: Hyllner
authors.forename: Björn
authors.initials: B
authors.lastname: Olsson
authors.forename: Peter
authors.initials: P
authors.lastname: Sartipy
authors.forename: Petter
authors.initials: P
authors.lastname: Björquist
mesh_terms: D000818:Animals; D002454:Cell Differentiation / Q000502:physiology*; D019070:Cell Lineage / Q000235:genetics*; D002478:Cells, Cultured; D053595:Embryonic Stem Cells / Q000166:cytology / Q000502:physiology*; D004707:Endoderm / Q000166:cytology* / Q000502:physiology; D020869:Gene Expression Profiling; D018507:Gene Expression Regulation, Developmental; D005819:Genetic Markers; D022781:Hepatocytes / Q000166:cytology* / Q000502:physiology; D006801:Humans; D020411:Oligonucleotide Array Sequence Analysis; D014158:Transcription, Genetic; D015854:Up-Regulation
chemical_list: D005819:Genetic Markers