Donor-dependent variations in hepatic differentiation from human-induced pluripotent stem cells

Abstract

Hepatocytes generated from human induced pluripotent stem cells (hiPSCs) are unprecedented resources for pharmaceuticals and cell therapy. However, the in vitro directed differentiation of human pluripotent stem cells into mature hepatocytes remains challenging. Little attention has so far been paid to variations among hiPSC lines in terms of their hepatic differentiation. In the current study, we developed an improved hepatic differentiation protocol and compared 28 hiPSC lines originated from various somatic cells and derived using retroviruses, Sendai viruses, or episomal plasmids. This comparison indicated that the origins, but not the derivation methods, may be a major determinant of variation in hepatic differentiation. The hiPSC clones derived from peripheral blood cells consistently showed good differentiation efficiency, whereas many hiPSC clones from adult dermal fibroblasts showed poor differentiation. However, when we compared hiPSCs from peripheral blood and dermal fibroblasts from the same individuals, we found that variations in hepatic differentiation were largely attributable to donor differences, rather than to the types of the original cells. These data underscore the importance of donor differences when comparing the differentiation propensities of hiPSC clones.

Tankyrase inhibition promotes a stable human naïve pluripotent state with improved functionality

“…Previous studies suggested that reprogrammingassociated errors of retention of donor cell-specific epigenetic memory bias the differentiation potency of hiPSCs toward some lineages (Kim et al, 2010b(Kim et al, , 2011Polo et al, 2010). However, other studies did not confirm such correlations, or alternatively suggested that donor-specific genetic variability affecting lineage-primed gene expression might play a more dominant role (Ohi et al, 2011;Hu et al, 2011;Kajiwara et al, 2012;Kyttälä et al, 2016).…”

Induction of pluripotency in human somatic cells via a transient state resembling primitive streak-like mesendoderm

“…EN differentiation. EN differentiation was performed as described previously, with slight modification 36 . The single-cell suspensions of human pluripotent stem cells were plated onto Matrigel-coated plates in RPMI1640 (Life Technologies) containing 2% B27, 100 ng ml À 1 Activin A, 3 mM CHIR99021 and 0.5% Pen/Strep.…”

Multicolor Staining of Globin Subtypes Reveals Impaired Globin Switching During Erythropoiesis in Human Pluripotent Stem Cells

“…The hiPSC technology could also contribute to production of rare types of erythrocytes-for example, D2, Fy(a2b2), Di(b2), and Jr(a2) [5]. However, it is well known that hiPSC clones are heterogeneous with respect to their differentiation potential [6]. Consequently, the ability to distinguish clones suitable for in vitro erythrocyte production from among identical donor-derived hiPSC clones is crucial.…”