A single-chain derivative of an integrin-activating antibody potentiates organoid growth in Matrigel and collagen hydrogels
Wim B. M. de Lau, Joost J.A.P.M. Wijnakker, Gijs J. F. van Son, Daniel Krueger, Daisong Wang, Matthijs S. Abendroth, Robin Schreurs, Claudia Y. Janda, Fenna L. H. van Rijt, Benjamin Chalopin, Emma Bokobza, Johan H. van Es, Timothy A. Springer, Arnoud Sonnenberg, Hans Clevers
Abstract
Current methods of culturing human epithelial organoids from adult stem cells may not be compatible with clinical applications as they rely on xenogeneic, chemically undefined or non-standardized components such as the basement membrane extract Matrigel. Matrigel provides a source of extracellular matrix molecules, including laminins and collagen IV, which interact with β1 integrins expressed on organoid cells. Here we describe a single-chain (sc) version of antibody TS2/16 that allosterically activates integrin β1 function in organoids. The addition of monomeric scTS2/16 to organoid medium results in up to a fivefold increase in the yield of all gastrointestinal organoids grown in Matrigel. Moreover, scTS2/16 supports a six- to sevenfold increase in the yield of these organoids when cultured in collagen I hydrogels, both in 3D and 2D. Collagen I is well defined, available in clinical-grade formulations and, when combined with scTS2/16, may support the clinical application of epithelial organoids derived from gastrointestinal tissues and other epithelial sources.