Systematic optimization of culture media for maintenance of human induced pluripotent stem cells using the response surface methodology

Abstract

The application of human induced pluripotent stem cells (hiPSCs) provides tremendous opportunities in cell therapy. However, culturing these cells faces many practical challenges, including costs associated with cell culture media and the optimization of cell culture conditions. Providing an optimized culture platform for hiPSCs to maintain pluripotency and self-renewal and generate cost-effective and robust therapeutics is an immediate requirement. This study used the design of experiments and the response surface methodology, a powerful statistical tool, to generate empirical models for predicting the optimal culture conditions of the hiPSCs. Pluripotency and cell proliferation were applied as read-outs to determine the optimal concentration of basic fibroblast growth factor (bFGF) and cell density. One model was defined to predict pluripotency and cell proliferation in terms of the predictor variables of the bFGF concentration and cell seeding density. Predicted culture conditions to maximize maintaining cell pluripotency were successfully validated. The present study's findings provide a novel approach that can potentially allow controllable hiPSC culture routine in translational research.

Highlights

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  Application of response surface methodology optimized bFGF and cell density for hiPSC culture.

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  The RSM approach minimizes experiments, offering a cost-effective and controllable hiPSC culture.

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  This Study identified optimal conditions (bFGF 111 ng/ml, 70,000 cells/cm ²) for hiPSC expansion.

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  Predicted conditions (bFGF 130 ng/ml, 70,000 cells/cm ²) maintained pluripotency.

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  Optimal conditions enhance pluripotency marker expression, critical for translational research.