Massaro Angelo, Villegas Novoa Cecilia, Wang Yuli, Allbritton Nancy L
Department of Bioengineering, University of Washington, Seattle, WA, United States.
Front Bioeng Biotechnol. 2024 Dec 16;12:1506976. doi: 10.3389/fbioe.2024.1506976. eCollection 2024.
Colonic epithelium is situated above a layer of fibroblasts that provide supportive factors for stem cells at the crypt base and promote differentiation of cells in the upper crypt and luminal surface. To study the fibroblast-epithelial cell interactions, an crypt model was formed on a shaped collagen scaffold with primary epithelial cells growing above a layer of primary colonic fibroblasts. The crypts possessed a basal stem cell niche populated with proliferative cells and a differentiated, nondividing cell zone at the luminal crypt end. The presence of fibroblasts enhanced cell differentiation and accelerated the rate at which a high resistance epithelial cell layer formed relative to cultures without fibroblasts. The fibroblasts modulated cell proliferation within crypts increasing the number of crypts populated with proliferative cells but decreasing the total number of proliferative cells in each crypt. Bulk-RNA sequencing revealed 41 genes that were significantly upregulated and 190 genes that were significantly downregulated in cocultured epithelium relative to epithelium cultured without fibroblasts. This epithelium-fibroblast crypt model suggests bidirectional communication between the two cell types and has the potential to serve as a model to investigate fibroblast-epithelial cell interactions in health and disease.
结肠上皮位于一层成纤维细胞之上，这些成纤维细胞为隐窝底部的干细胞提供支持因子，并促进隐窝上部和管腔表面细胞的分化。为了研究成纤维细胞与上皮细胞之间的相互作用，在一个成型的胶原蛋白支架上形成了一个隐窝模型，原代上皮细胞生长在一层原代结肠成纤维细胞之上。这些隐窝拥有一个充满增殖细胞的基底干细胞龛，以及在隐窝管腔末端的一个分化的、不分裂的细胞区。与没有成纤维细胞的培养物相比，成纤维细胞的存在增强了细胞分化，并加快了高抗性上皮细胞层形成的速度。成纤维细胞调节隐窝内的细胞增殖，增加了充满增殖细胞的隐窝数量，但减少了每个隐窝中增殖细胞的总数。批量RNA测序显示，与没有成纤维细胞培养的上皮相比，共培养上皮中有41个基因显著上调，190个基因显著下调。这种上皮-成纤维细胞隐窝模型表明两种细胞类型之间存在双向通讯，并且有潜力作为一个模型来研究健康和疾病状态下成纤维细胞与上皮细胞之间的相互作用。
