Splitting hESC/hiPSC lines with EDTA in feeder-free conditions

Abstract

Individualization leads to severe cell death in human pluripotent stem cells (hPSCs), including human embryonic stem cells (hESCs) and human induced pluripotent stem cells (hiPSCs). This phenomenon leads to difficulties in handling cells in applications such as passaging, cryopreservation and transfection, and the treatment with ROCK inhibitors has been found to effectively improve cell survival. However, enzymatic dissociation and ROCK inhibitor treatment could bring potential complication in cell maintenance. An enzyme-free dissociation method was developed to passage stem cells and achieve robust cell survival in dependent of drug treatment.