Elimination of undifferentiated human embryonic stem cells by cardiac glycosides

Abstract

An important safety concern in the use of human pluripotent stem cells (hPSCs) is tumorigenic risk, because these cells can form teratomas after an in vivo injection at ectopic sites. Several thousands of undifferentiated hPSCs are sufficient to induce teratomas in a mouse model. Thus, it is critical to remove all residue-undifferentiated hPSCs that have teratoma potential before the clinical application of hPSC-derived cells. In this study, our data demonstrated the cytotoxic effects of cardiac glycosides, such as digoxin, lanatoside C, bufalin, and proscillaridin A, in human embryonic stem cells (hESCs). This phenomenon was not observed in human bone marrow mesenchymal stem cells (hBMMSCs). Most importantly, digoxin and lanatoside C did not affect the stem cells’ differentiation ability. Consistently, the viability of the hESC-derived MSCs, neurons, and endothelium cells was not affected by the digoxin and lanatoside C treatment. Furthermore, the in vivo experiments demonstrated that digoxin and lanatoside C prevented teratoma formation. To the best of our knowledge, this study is the first to describe the cytotoxicity and tumor prevention effects of cardiac glycosides in hESCs. Digoxin and lanatoside C are also the first FDA-approved drugs that demonstrated cytotoxicity in undifferentiated hESCs.

Insulin Stimulates PI3K/AKT and Cell Adhesion to Promote the Survival of Individualized Human Embryonic Stem Cells

“…When hPSCs are differentiated to specific cell types, it is often necessary to remove undifferentiated hPSCs from the cell population. Conventionally, small chemicals are used to kill hPSCs . Here, we examined whether insulin‐free passaging can be used to eliminate undifferentiated cells.…”

A novel method to reconstruct epithelial tissue using high-purity keratinocyte lineage cells induced from human embryonic stem cells

“…In transplantation studies, hESCs were capable of differentiating within biological scaffolds to form a variety of cell types with the characteristics of normal tissues. However, the cells also formed teratomas [28], which limits their use in transplantation [29]. The biological scaffold material provides a good environment for adhesion, growth, migration, proliferation, and differentiation.…”

Peer Review #1 of "Enhanced genome editing in human iPSCs with CRISPR-CAS9 by co-targeting ATP1a1 (v0.1)"

“…Cardiac glycosides have been used to treat heart failure by targeting ATP1A1, a subunit of Na + /K + -ATPase (McDonough et al 2002;Smith 1984). With exposure to relatively high concentrations of such drugs, cell viability is reduced via the accumulation of intracellular Ca 2+ levels (Belusa et al 2002;Lin et al 2017). The binding site of cardiac glycosides on ATP1A1 has been identified, and N-terminal amino-acid substitution of ATP1A1 encoded by exon4 (Q118R and N129D) is sufficient to confer drug resistance by preventing the binding of the cardiac glycosides (Treschow et al 2007).…”