Contribution of Hepatic Lineage Stage‐Specific Donor Memory to the Differential Potential of Induced Mouse Pluripotent Stem Cells

Abstract

Recent studies suggested that induced pluripotent stem cells (iPSCs) retain a residual donor cell gene expression which may impact their capacity to differentiate into cell of origin. Here we addressed a contribution of a lineage stage-specific donor cell memory in modulating the functional properties of iPSCs. iPSCs were generated from hepatic lineage cells at an early (hepatoblast-derived, HB-iPSCs) and end stage (adult hepatocyte, AH-iPSCs) of hepatocyte differentiation as well as from mouse fetal fibroblasts (MEF-iPSCs) using a lentiviral vector encoding four pluripotency-inducing factors Oct4, Sox2, Klf4, and c-Myc. All resulting iPS cell lines acquired iPSCs phenotype as judged by the accepted criteria including morphology, expression of pluripotency markers, silencing of transducing factors, capacity of multilineage differentiation in teratoma assay and normal diploid karyotype. However, HB-iPSCs were more efficient in directed differentiation towards hepatocytic lineage as compared to AH-iPSCs, MEF-iPSCs or mESCs. Extensive comparative transcriptome analyses of the early passage iPSCs, donor cells and mESCs revealed that despite global similarities in gene expression patterns between generated iPSCs and mESCs, HB-iPSCs retained a transcriptional memory (7 up- and 17 down-regulated genes) typical of the original cells. Continuous passaging of HB-iPSCs erased most of these differences including a superior capacity for hepatic re-differentiation. These results suggest that retention of lineage stage-specific donor memory in iPSCs may facilitate differentiation into donor cell type. The identified gene set help to improve hepatic differentiation for therapeutic applications and contribute to the better understanding of liver development.

Specific fate decisions in adult hepatic progenitor cells driven by MET and EGFR signaling

“…Notably, signaling through EGFR failed to compensate for the lack of MET receptor in HPCs, while HGF stimulation of Egfr-null progenitor cells or MET restoration in MET mutant HPCs was sufficient to promote hepatocyte differentiation. These observations corroborate prior findings regarding the role of HGF/MET signaling in facilitating progenitor, embryonic stem cell, and induced pluripotent stem cell differentiation along hepatocyte lineage (Si-Tayeb et al 2010;Ishikawa et al 2012; Lee et al 2012). We further identify EGFR as a key player in cholangiocyte differentiation through a mechanism requiring NOTCH1.…”

Generation of Functional Mesenchymal Stem Cells from Different Induced Pluripotent Stem Cell Lines

“…Various studies have described the presence of the epigenetic memory of iPSC for their somatic tissue of origin that can influence the in vitro differentiation capacity of the iPSC [48][49][50]. This memory was, subsequently, lost through a continuous subculture of iPSC [48][49][50].…”

“…Various studies have described the presence of the epigenetic memory of iPSC for their somatic tissue of origin that can influence the in vitro differentiation capacity of the iPSC [48][49][50]. This memory was, subsequently, lost through a continuous subculture of iPSC [48][49][50]. More recent reports have, however, shown that iPSC retain the epigenetic memory of their tissue of origin even after an extensive passage [51][52][53][54], where the epigenetic memory skews the developmental potential of iPSC toward their previous state of differentiation [51][52][53][54].…”

Cell sources forin vitrohuman liver cell culture models

“…137 Thus, the derivation of hiPSC lines from hepatic cells could lead to enhanced hepatic differentiation, as already reported for mouse iPSC. 138 However, the so-called epigenetic memory is also influenced by the culture conditions, e.g. the number of passages.…”