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 -