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首页> 外文期刊>Journal of Cellular Physiology >Use of a Collagen Membrane to Enhance the Survival of Primary Intestinal Epithelial Cells
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Use of a Collagen Membrane to Enhance the Survival of Primary Intestinal Epithelial Cells

机译：使用胶原蛋白膜提高生存原发性肠道上皮细胞

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摘要

Intestinal epithelial cell culture is important for biological, functional, and immunological studies. Since enterocytes have a short in vivo life span due to anoikis, we aimed to establish a novel and reproducible method to prolong the survival of mouse and human cells. Cells were isolated following a standard procedure, and cultured on ordered-cow's collagen membranes. A prolonged cell life span was achieved; cells covered the complete surface of bio-membranes and showed a classical enterocyte morphology with high expression of enzymes supporting the possibility of cryopreservation. Apoptosis was dramatically reduced and cultured enterocytes expressed cytokeratin and LGR5 (low frequency). Cells exposed to LPS or flagellin showed the induction of TLR4 and TLR5 expression and a functional phenotype upon exposure to the probiotic Bifidobacterium bifidum or the pathogenic Clostridium difficile. The secretion of the homeostatic (IL-25 and TSLP), inhibitory (IL-10 and TGF-beta), or pro-inflammatory mediators (IL-1 beta and TNF) were induced. In conclusion, this novel protocol using cow's collagen-ordered membrane provides a simple and reproducible method to maintain intestinal epithelial cells functional for cell-microorganism interaction studies and stem cell expansion. (C) 2016 Wiley Periodicals, Inc.
机译：肠上皮细胞培养是很重要的生物、功能以及免疫研究。由于女性的寿命,我们旨在建立一个小说和可再生的方法来延长老鼠和人类细胞的生存。孤立的标准程序后,培养ordered-cow胶原蛋白膜。延长细胞寿命得以实现;完整的bio-membranes表面和覆盖显示一个经典肠上皮细胞形态酶的高表达支持低温贮藏的可能性。和培养肠上皮细胞明显减少表达细胞角蛋白和LGR5就(低频)。细胞暴露于有限合伙人或鞭毛蛋白显示感应TLR4和TLR5表达式功能表型在接触益生菌双歧杆菌bifidum或梭状芽胞杆菌致病。稳态(IL-25和TSLP)的抑制(il - 10和鉴定及),或促炎症介质(il - 1β和TNF)诱导。结论,这部小说用牛的协议collagen-ordered膜提供了一个简单的和可再生的方法维持肠道上皮细胞功能cell-microorganism交互研究和干细胞细胞扩张。

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