top of page


Large-scale optimisation of human pluripotent stem cell culture conditions

V Pisupati(1) S Srinivasaraghavan(1) S A Mahmoud(1) I Mali(1) M Perez-Alcantara(3) R Barker(1) I Barbaric(2) D Chiarugi(4) F T Merkle(1)

1:University of Cambridge; 2:The University of Sheffield; 3:Wellcome Sanger Institute; 4:Max Planck Institute for Human Cognitive and Brain Sciences

During their time in culture, human pluripotent stem cells (hPSCs) recurrently acquire mutations that compromise their utility for disease modelling and regenerative medicine, including in  cancer-associated genes that are of particular concern for human cell transplantation therapies. The frequency with which these mutations recur suggest that hPSC culture conditions are sub-optimal, but the wide array of different growth conditions that are used today have complicated efforts to compare them directly and identify those that reduce selective pressures experienced by hPSCs. Here, we report a novel method for directly comparing the effects of dozens of different media, substrates, passaging methods, on the absolute and relative growth rates of hPSCs to indirectly quantify selective pressures. Having identified those conditions that minimise selective pressures, we further tested their ability to suppress the growth advantages of p53 mutant cell lines to suggest a set of more optimal growth conditions. Together, we hope these studies will promote the standardisation, reproducibility, and safety of hPSC-based cell therapies.

bottom of page