Bergenheim Fredrik, Fregni Giulia, Buchanan Cara Field, Riis Lene Buhl, Heulot Mathieu, Touati Jeremy, Seidelin Jakob Benedict, Rizzi Simone Carlo, Nielsen Ole Haagen
Dept. of Gastroenterology, Herlev Hospital, University of Copenhagen, 2730 Herlev, Denmark.
QGel SA, Innovation Park, EPFL Building G, 1015 Lausanne, Switzerland.
Biomaterials. 2020 Dec;262:120248. doi: 10.1016/j.biomaterials.2020.120248. Epub 2020 Aug 19.
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.
肠道类器官有着广泛的研究和生物医学应用，如疾病建模、药物测试和再生医学。然而，向临床应用的转变在一定程度上受到对动物肿瘤来源的基底膜提取物（BMEs）的依赖的阻碍，这些提取物定义不明确，且由于其来源和批次间的变异性而不适合监管批准。为了克服这些限制并实现临床转化，我们测试了使用一种完全确定的水凝胶基质QGel CN99，直接从人类结肠活检组织中建立和扩增肠道类器官。我们实现了高效的从头建立、扩增和类器官维持，同时还证明了持续的遗传稳定性。此外，我们能够保留干性和分化能力，其转录组谱类似于正常结肠上皮。所有数据都证明与在BME基准基质胶中培养的类器官相当。完全绕过BMEs使用的完全确定的水凝胶的应用，将极大地提高类器官研究的可重复性和可扩展性，同时也推动个性化医学和基于干细胞的再生疗法中的转化应用。
