TY - JOUR

T1 - A fully defined 3D matrix for ex vivo expansion of human colonic organoids from biopsy tissue

AU - Bergenheim, Fredrik

AU - Fregni, Giulia

AU - Buchanan, Cara Field

AU - Riis, Lene Buhl

AU - Heulot, Mathieu

AU - Touati, Jeremy

AU - Seidelin, Jakob Benedict

AU - Rizzi, Simone Carlo

AU - Nielsen, Ole Haagen

PY - 2020

Y1 - 2020

N2 - Intestinal organoids have widespread research and biomedical applications, such as disease modeling, drug testing and regenerative medicine. However, the transition towards clinical use has in part been hampered by the dependency on animal tumor-derived basement membrane extracts (BMEs), which are poorly defined and ill-suited for regulatory approval due to their origin and batch-to-batch variability. In order to overcome these limitations, and to enable clinical translation, we tested the use of a fully defined hydrogel matrix, QGel CN99, to establish and expand intestinal organoids directly from human colonic biopsies. We achieved efficient de novo establishment, expansion and organoid maintenance, while also demonstrating sustained genetic stability. Additionally, we were able to preserve stemness and differentiation capacity, with transcriptomic profiles resembling normal colonic epithelium. All data proved comparable to organoids cultured in the BME-benchmark Matrigel. The application of a fully defined hydrogel, completely bypassing the use of BMEs, will drastically improve the reproducibility and scalability of organoid studies, but also advance translational applications in personalized medicine and stem cell-based regenerative therapies.

AB - Intestinal organoids have widespread research and biomedical applications, such as disease modeling, drug testing and regenerative medicine. However, the transition towards clinical use has in part been hampered by the dependency on animal tumor-derived basement membrane extracts (BMEs), which are poorly defined and ill-suited for regulatory approval due to their origin and batch-to-batch variability. In order to overcome these limitations, and to enable clinical translation, we tested the use of a fully defined hydrogel matrix, QGel CN99, to establish and expand intestinal organoids directly from human colonic biopsies. We achieved efficient de novo establishment, expansion and organoid maintenance, while also demonstrating sustained genetic stability. Additionally, we were able to preserve stemness and differentiation capacity, with transcriptomic profiles resembling normal colonic epithelium. All data proved comparable to organoids cultured in the BME-benchmark Matrigel. The application of a fully defined hydrogel, completely bypassing the use of BMEs, will drastically improve the reproducibility and scalability of organoid studies, but also advance translational applications in personalized medicine and stem cell-based regenerative therapies.

KW - Cell differentiation

KW - Extracellular matrix

KW - Human colonic organoids

KW - Hydrogel

KW - Intestinal stem cells

U2 - 10.1016/j.biomaterials.2020.120248

DO - 10.1016/j.biomaterials.2020.120248

M3 - Journal article

C2 - 32891909

AN - SCOPUS:85090159010

VL - 262

JO - Biomaterials

JF - Biomaterials

SN - 0142-9612

M1 - 120248

ER -