Hepatic population derived from human pluripotent stem cells is effectively increased by selective removal of undifferentiated stem cells using YM155

Abstract

BackgroundPluripotent stem cells (PSCs) such as embryonic stem cells and induced pluripotent stem cells are promising target cells for cell regenerative medicine together with recently advanced technology of in-vitro differentiation. However, residual undifferentiated stem cells (USCs) during in-vitro differentiation are considered a potential risk for development of cancer cells and nonspecific lineage cell types. In this study we observed that USCs still exist during hepatic differentiation, consequently resulting in poor quality of the hepatic population and forming teratoma in vivo. Therefore, we hypothesized that effectively removing USCs from in-vitro differentiation could improve the quality of the hepatic population and guarantee safety from risk of teratoma formation.MethodsHuman PSCs were differentiated to hepatocytes via four steps. YM155, a known BIRC5 inhibitor, was applied for removing the residual USCs on the hepatic differentiation. After YM155 treatment, hepatocyte development was evaluated by measuring gene expression, immunostaining and hepatic functions at each stage of differentiation, and forming teratomas were confirmed by cell transplantation with or without YM155.ResultsThe selected concentrations of YM155 removed USCs (NANOG+ and OCT4+) in a dose-dependent manner. As a result, expression of endodermal markers (SOX17, FOXA2 and CXCR4) at stage II of differentiation and hepatic markers (ALB, AFP and HNF4A) at stage III was up-regulated by YM155 treatment as well as the hepatic population (ALB+), and functions (ALB/urea secretion and CYP450 enzyme activity) were enhanced at the final stage of differentiation (stage IV). Furthermore, we demonstrated that NANOG and OCT4 expression remaining until stage III (day 15 of differentiation) completely disappeared when treated with YM155 and teratoma formation was effectively prevented by YM155 pretreatment in the in-vitro study.ConclusionsWe suggest that the removal of USCs using YM155 could improve the quantity and quality of induced hepatocytes and eliminate the potential risk of teratoma formation.Electronic supplementary materialThe online version of this article (doi:10.1186/s13287-017-0517-2) contains supplementary material, which is available to authorized users.

Prunellae Spica Extract Suppresses Teratoma Formation of Pluripotent Stem Cells through p53-Mediated Apoptosis

“…It has recently been reported that during hepatocyte differentiation, treatment with YM155, which is known to induce selective and complete cell death of undifferentiated hiPSCs, improved the quantity and quality of induced hepatocytes and consequently eliminated the possibility of teratoma formation [43]. Based on these observations, and our present findings, differentiation protocols can be devised to obtain safe cell sources with no risk of teratoma formation.…”

Scarless Enriched selection of Genome edited Human Pluripotent Stem Cells Using Induced Drug Resistance

“…By correlating the hPSC scores with the sensitivities of each cell line to each of 543 compounds, we identified YM155 as the most effective drug in cells with high hPSC scores (Fig. 1B); this implies that YM155 should be highly specific for hPSCs, as demonstrated previously (Bedel et al, 2017; Kang et al, 2017; Kim et al, 2017; Lee et al, 2013b). Based on correlation of drug and gene expression signature (Fig.…”

“…To address this danger, several approaches have been developed for selective removal of residual hPSCs (Jeong and Cha, 2017). For example, we previously reported that YM155, a survivin inhibitor developed as an anti-cancer drug, is selectively cytotoxic against undifferentiated hPSCs and inhibits teratoma formation (Lee et al, 2013b), and this finding has been reproduced in several independent studies (Bedel et al, 2017; Kang et al, 2017; Kim et al, 2017). At that time, however, the molecular mechanism underlying the high susceptibility of hPSCs to YM155 had not been fully elucidated.…”

“…Selective induction of cell death in hPSCs by YM155, which decreases teratoma formation (Bedel et al, 2017; Kang et al, 2017; Kim et al, 2017; Lee et al, 2013b), is associated with the DNA damage response (Lee et al, 2013b). Cytotoxicity of YM155 cannot be fully prevented by suppression of BIRC5 (which encodes survivin) (Sim et al, 2017).…”

Oncogenic STRAP Supports Hepatocellular Carcinoma Growth by Enhancing Wnt/β-Catenin Signaling

“…Con#23 Con#26 Conversely, STRAP loss increased most liver differentiation related genes, such as ALB, AFP, and HNF4A (35,36), in Huh6 and more obviously in Huh7 cells ( Supplementary Fig. S6B).…”