D-3 (phosphorpeptide) is an efficient, simple, and specific agent for eliminating iPSCs.

D-3 effectively inhibits the formation of teratomas by residual induced pluripotent stem cells (iPSCs) in a mouse model, as demonstrated in a tumorigenicity assay[1]. It significantly reduces the viability of 201B7 cells, with a half maximal inhibitory concentration (IC50) of 192.3 ± 57.4 μM[1]. The compound promotes an increase in the activation of proteins such as elF2a, p38, and p44/42 MAPK, which are known to respond to cellular stress. Additionally, D-3 treatment results in a higher incidence of apoptotic and dead cells as indicated by an increase in Annexin-V and SYTOX-positive cells[1]. Importantly, D-3 exhibits minimal effects on various non-iPSC cell types, including hepatocytes and neurons[1]. A Cell Viability Assay reveals that a concentration of 400 μM D-3 for 1-2 hours is sufficient to significantly decrease cell viability in six human iPSC lines and the human embryonic stem cell (ESC) line khES1, with over 99% viability loss in all iPSC lines and more than 95% in khES1[1].