Enhanced single-cell viability using 30Kc6 for efficient expansion of human induced pluripotent stem cells

Abstract

Human pluripotent stem cells (hPSCs), such as human induced pluripotent stem cells (hiPSCs) and human embryonic stem cells (hESCs), have been widely used in stem cell research. hPSCs form colonies in culture dishes and are passaged as clumps. If the cells are dissociated into single cells, epithelial structures are disrupted, and rapid apoptosis is induced. The low viability of single cells restricts the passaging and expansion of hPSCs. Here, in order to enhance the single cell viability of hPSCs, we adopted an apoptotic strategy using the 30 Kc6 gene which inhibits mitochondrial apoptosis. With the expression of the 30 Kc6 gene in hiPSCs, no issues related to pluripotency or differentiation occurred. The hiPSC-30 Kc6 showed higher viability after the induction of apoptosis compared with control hiPSCs. Furthermore, hiPSC-30 Kc6 formed more colonies when the cells were enzymatically dissociated into single cells. Taken together, this study demonstrates that the 30 Kc6 gene could facilitate the passage and expansion of hiPSCs, potentially preventing the dissociation-induced apoptosis of single hPSCs.