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bioRxivCell BiologyPreprint

A synthetic xeno-free extracellular matrix surrogate supports robust human induced pluripotent stem cell maintenance and multilineage differentiation

Constantin BergeriD, Linus WioraiD, Terri L. HolmesiD, Fabiola Walz, Clemens M. SauteriD, Luisa Baraban, Julian Schwebler, Geraldine Beer, Sonja Lauke, Jana Keller, Melanie Kraft, Yvonne SchellingiD, Ka Cheung, Elina NürnbergiD, Sarah Al Muthafer, Sandra Segeletz, Christina Neser, James G.W. SmithiD, Stefan HauseriD, Daniela ZdziebloiD

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Abstract

Animal-derived surface coatings present the gold standard in stem cell culture. However, they come with major drawbacks such as lot-to-lot inconsistency and an ethically problematic production. Approaches to develop synthetic coating alternatives have largely failed to recapitulate biological functions essential to PSC-self renewal and differentiation. In this study, we have investigated the suitability of a fully synthetic, chemically defined extracellular matrix-mimicking coating named BIOINSTRUCT for hiPSC culture. BIOINSTRUCT was employed during maintenance culture of IMR90-4 cells, as well as directed differentiation of HIHCNi008-A, ACTN2-GFP, and UKKI-011-A cells. BIOINSTRUCT preserved hiPSC characteristics over the experimental period of 10 passages similar to Matrigel. Spontaneous differentiation resulted in the formation of ectodermal, mesodermal and endodermal derivatives, confirming the retention of pluripotency. Directed differentiation on BIOINSTRUCT enabled the generation of neuronal, cardiac and pancreatic endocrine cells. Altogether, our data underscore BIOINSTRUCT as a promising alternative to animal-derived extracellular matrix coatings.

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