A fully defined 3D matrix for ex vivo expansion of human colonic organoids from biopsy tissue. (December 2020)

• Record Type: Journal Article
• Authors: Bergenheim, Fredrik; Fregni, Giulia; Buchanan, Cara Field; Riis, Lene Buhl; Heulot, Mathieu; Touati, Jeremy; Seidelin, Jakob Benedict; Rizzi, Simone Carlo; Nielsen, Ole Haagen
• Is Part Of: Biomaterials. Volume 262(2020)
• Publication Date: 2020-12
• Subjects: Cell differentiation -- Extracellular matrix -- Human colonic organoids -- Hydrogel -- Intestinal stem cells; Biomedical materials -- Periodicals; Biocompatible Materials -- Periodicals; Biomatériaux -- Périodiques; 610.28
• DOI: 10.1016/j.biomaterials.2020.120248
• Languages: English
• ISSNs: 0142-9612

Abstract

Abstract: 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. Highlights: QGel CN99 is a novel fully defined hydrogel. We show efficient establishment of human colon organoids from fresh biopsies in CN99. Cell expansion, stemness and differentiation capacity are comparable to Matrigel. Transcriptomic profiles of cultured organoids resemble corresponding healthy tissue. The use of a basal membrane extract (e.g., Matrigel) can be completely bypassed.