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.

肠类器官具有广泛的研究和生物医学应用，例如疾病建模，药物测试和再生医学。但是，对临床应用的过渡部分地受到了对动物肿瘤来源的基底膜提取物（BMEs）的依赖的阻碍，这些提取物的定义不明确，并且由于其来源和批次之间的差异性而不适用于监管部门的批准。为了克服这些局限性并实现临床翻译，我们测试了使用完全定义的水凝胶基质QGel CN99来直接从人类结肠活检组织中建立和扩展肠道类器官。我们实现了从头开始的高效建立，扩展和类器官维持，同时也证明了持续的遗传稳定性。此外，我们能够保留茎干和分化能力，其转录组特征类似于正常结肠上皮。所有数据证明与在BME基准Matrigel中培养的类器官具有可比性。完全绕过BME的使用，完全定义的水凝胶的应用将大大改善类器官研究的可重复性和可扩展性，而且还将促进翻译在个性化医学和基于干细胞的再生疗法中的应用。