title: Highly Synchronized Expression of Lineage-Specific Genes during In Vitro Hepatic Differentiation of Human Pluripotent Stem Cell Lines.
abstract: Human pluripotent stem cells- (hPSCs-) derived hepatocytes have the potential to replace many hepatic models in drug discovery and provide a cell source for regenerative medicine applications. However, the generation of fully functional hPSC-derived hepatocytes is still a challenge. Towards gaining better understanding of the differentiation and maturation process, we employed a standardized protocol to differentiate six hPSC lines into hepatocytes and investigated the synchronicity of the hPSC lines by applying RT-qPCR to assess the expression of lineage-specific genes (OCT4, NANOG, T, SOX17, CXCR4, CER1, HHEX, TBX3, PROX1, HNF6, AFP, HNF4a, KRT18, ALB, AAT, and CYP3A4) which serve as markers for different stages during liver development. The data was evaluated using correlation and clustering analysis, demonstrating that the expression of these markers is highly synchronized and correlated well across all cell lines. The analysis also revealed a distribution of the markers in groups reflecting the developmental stages of hepatocytes. Functional analysis of the differentiated cells further confirmed their hepatic phenotype. Taken together, these results demonstrate, on the molecular level, the highly synchronized differentiation pattern across multiple hPSC lines. Moreover, this study provides additional understanding for future efforts to improve the functionality of hPSC-derived hepatocytes and thereby increase the value of related models.
authors.affiliation: School of Bioscience, Systems Biology Research Center, University of Skövde, 541 28 Skövde, Sweden; Institute of Biomedicine, Department of Clinical Chemistry and Transfusion Medicine, Sahlgrenska Academy, University of Gothenburg, 413 45 Gothenburg, Sweden.
authors.forename: Nidal
authors.initials: N
authors.lastname: Ghosheh
authors.affiliation: School of Bioscience, Systems Biology Research Center, University of Skövde, 541 28 Skövde, Sweden.
authors.forename: Björn
authors.initials: B
authors.lastname: Olsson
authors.affiliation: Takara Bio Europe AB, Arvid Wallgrens Backe 20, 413 46 Gothenburg, Sweden.
authors.forename: Josefina
authors.initials: J
authors.lastname: Edsbagge
authors.affiliation: Takara Bio Europe AB, Arvid Wallgrens Backe 20, 413 46 Gothenburg, Sweden.
authors.forename: Barbara
authors.initials: B
authors.lastname: Küppers-Munther
authors.affiliation: Takara Bio Europe AB, Arvid Wallgrens Backe 20, 413 46 Gothenburg, Sweden.
authors.forename: Mariska
authors.initials: M
authors.lastname: Van Giezen
authors.affiliation: School of Bioscience, Systems Biology Research Center, University of Skövde, 541 28 Skövde, Sweden.
authors.forename: Annika
authors.initials: A
authors.lastname: Asplund
authors.affiliation: AstraZeneca R&D, CVMD DMPK, 431 83 Mölndal, Sweden; Department of Physiology and Pharmacology, Section of Pharmacogenetics, Karolinska Institutet, 171 77 Stockholm, Sweden.
authors.forename: Tommy B
authors.initials: TB
authors.lastname: Andersson
authors.affiliation: NovaHep AB, Arvid Wallgrens Backe 20, 413 46 Gothenburg, Sweden.
authors.forename: Petter
authors.initials: P
authors.lastname: Björquist
authors.affiliation: Sahlgrenska Cancer Center, Department of Pathology, Institute of Biomedicine, Sahlgrenska Academy, University of Gothenburg, 405 30 Gothenburg, Sweden.
authors.forename: Helena
authors.initials: H
authors.lastname: Carén
authors.affiliation: Institute of Biomedicine, Department of Clinical Chemistry and Transfusion Medicine, Sahlgrenska Academy, University of Gothenburg, 413 45 Gothenburg, Sweden.
authors.forename: Stina
authors.initials: S
authors.lastname: Simonsson
authors.affiliation: School of Bioscience, Systems Biology Research Center, University of Skövde, 541 28 Skövde, Sweden; AstraZeneca R&D, GMD CVMD GMed, 431 83 Mölndal, Sweden.
authors.forename: Peter
authors.initials: P
authors.lastname: Sartipy
authors.affiliation: School of Bioscience, Systems Biology Research Center, University of Skövde, 541 28 Skövde, Sweden.
authors.forename: Jane
authors.initials: J
authors.lastname: Synnergren