Kakar Sachin, Derouet Mathieu F, Zhang Liyue, Gillis Connor J, Larsen Frederikke, Paul Arghya, Flynn Lauren E, Asfaha Samuel
Department of Pathology and Laboratory Medicine, Schulich School of Medicine and Dentistry, The University of Western Ontario, London, Ontario, Canada.
Department of Medicine, Schulich School of Medicine and Dentistry, The University of Western Ontario, London, Ontario, Canada.
PLoS One. 2025 Aug 6;20(8):e0328898. doi: 10.1371/journal.pone.0328898. eCollection 2025.
The generation of a tissue-specific intestinal hydrogel derived from the native intestine has the potential to support and promote the growth of intestinal organoids. In this study, we aimed to develop hydrogels derived exclusively from intestinal extracellular matrix (ECM) or composites comprised of intestinal ECM combined with alginate that allow for greater tuning of the hydrogel properties. A novel mouse intestinal decellularization protocol was developed and the ECM characterized. Our analyses demonstrate that our protocol effectively removed cellular and nuclear content while preserving key ECM components including collagens, glycosaminoglycans, fibronectin and laminin. When the decellularized small intestine (DSI) was used to generate hydrogels, the resulting ECM showed bioactivity as demonstrated by metabolic and pro-proliferative effects on NIH 3T3 murine fibroblasts. Importantly, our novel DSI hydrogels also supported murine intestinal and colonic organoid growth similar to Matrigel® controls. These studies demonstrate that murine tissue-specific DSI hydrogels can provide a supportive environment for the culture of intestinal and colonic organoids in vitro.
源自天然肠道的组织特异性肠道水凝胶的生成具有支持和促进肠道类器官生长的潜力。在本研究中，我们旨在开发仅由肠道细胞外基质（ECM）衍生的水凝胶或由肠道ECM与藻酸盐组成的复合材料，从而能够更好地调节水凝胶的特性。我们开发了一种新型的小鼠肠道去细胞化方案并对ECM进行了表征。我们的分析表明，我们的方案有效地去除了细胞和细胞核内容物，同时保留了包括胶原蛋白、糖胺聚糖、纤连蛋白和层粘连蛋白在内的关键ECM成分。当使用去细胞化小肠（DSI）来生成水凝胶时，所得的ECM表现出生物活性，这通过对NIH 3T3小鼠成纤维细胞的代谢和促增殖作用得以证明。重要的是，我们新型的DSI水凝胶也支持小鼠肠道和结肠类器官的生长，类似于基质胶®对照。这些研究表明，小鼠组织特异性DSI水凝胶可为体外培养肠道和结肠类器官提供支持性环境。
