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Optimization of the nutritional environment for differentiation of human‐induced pluripotent stem cells using design of experiments—A proof of concept

Esteban, PP; Patel, H; Veraitch, F; Khalife, R; (2021) Optimization of the nutritional environment for differentiation of human‐induced pluripotent stem cells using design of experiments—A proof of concept. Biotechnology Progress , Article e3143. 10.1002/btpr.3143. (In press). Green open access

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Abstract

The utilization of human‐induced pluripotent stem cells (hiPSCs) in cell therapy has a tremendous potential but faces many practical challenges, including costs associated with cell culture media and growth factors. There is an immediate need to establish an optimized culture platform to direct the differentiation of hiPSCs into germ layers in a defined nutritional microenvironment to generate cost‐effective and robust therapeutics. The aim of this study was to identify the optimal nutritional environment by mimicking the in vivo concentrations of three key factors (glucose, pyruvate, and oxygen) during the spontaneous differentiation of hiPSCs derived from cord blood, which greatly differ from the in vitro expansion and differentiation scenarios. Moreover, we hypothesized that the high glucose, pyruvate, and oxygen concentrations found in typical growth media could inhibit the differentiation of certain lineages. A design of experiments was used to investigate the interaction between these three variables during the spontaneous differentiation of hiPSCs. We found that lower oxygen and glucose concentrations enhance the expression of mesodermal (Brachyury, KIF1A) and ectodermal (Nestin, β‐Tubulin) markers. Our findings present a novel approach for efficient directed differentiation of hiPSCs through the manipulation of media components while simultaneously avoiding the usage of growth factors thus reducing costs.

Type: Article
Title: Optimization of the nutritional environment for differentiation of human‐induced pluripotent stem cells using design of experiments—A proof of concept
Location: United States
Open access status: An open access version is available from UCL Discovery
DOI: 10.1002/btpr.3143
Publisher version: https://doi.org/10.1002/btpr.3143
Language: English
Additional information: Copyright © 2021 The Authors. Biotechnology Progress published by Wiley Periodicals LLC on behalf of American Institute of Chemical Engineers. This is an open access article under the terms of the Creative Commons Attribution‐NonCommercial‐NoDerivs License (http://creativecommons.org/licenses/by-nc-nd/4.0/), which permits use and distribution in any medium, provided the original work is properly cited, the use is non‐commercial and no modifications or adaptations are made.
Keywords: Design of Experiments, Germ Layer Differentiation, Human Induced Pluripotent Stem Cells
UCL classification: UCL
UCL > Provost and Vice Provost Offices > UCL BEAMS
UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Engineering Science
UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Engineering Science > Dept of Biochemical Engineering
URI: https://discovery.ucl.ac.uk/id/eprint/10124886
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