Piper Hannah K, Wiese Mitchell J, Poss Mitchell J
Kemin Industries, Inc, Des Moines, IA, USA.
Sci Rep. 2025 Jul 22;15(1):26652. doi: 10.1038/s41598-025-12386-9.
Primary intestinal epithelial cells are a powerful tool to study tight junction protein function and barrier integrity of epithelial cells. However, primary cells are costly, difficult to isolate, have a finite ability to replicate, and require the usage of live tissue. An alternative to primary cells is immortalized cell lines, which are cost effective, easily cultured, stored, and maintain the physiology of primary cells. Several immortalized lines are commercially available, yet there remains a need for cell lines for livestock research, specifically bovine. Here we report an immortalized bovine intestinal epithelial cell (IBEC) line was established by transfection with exogenous human telomerase reverse transcriptase (hTERT). Primary bovine intestinal epithelial cells (PBECs) were isolated from a 2-day old mixed breed calf. PBECs were transfected with hTERT to create an IBEC line. The transfected cells were positive for intestinal epithelial markers while testing negative for fibroblast markers. There was no significant difference in IBEC morphology or physiology post immortalization. In Trans-Epithelial Electrical Resistance (TEER assays), PBECs and IBECs behaved similarly to intestinal porcine enterocyte cells (IPEC-J2) under treatment of Iipopolysaccharide and calcium butyrate. This newly-developed cell line will be an essential and useful tool for in vitro bovine and ruminant intestinal research.
原代肠上皮细胞是研究紧密连接蛋白功能和上皮细胞屏障完整性的有力工具。然而，原代细胞成本高昂、分离困难、复制能力有限，且需要使用活体组织。原代细胞的替代物是永生化细胞系，其具有成本效益、易于培养、储存，并能维持原代细胞的生理特性。有几种永生化细胞系可通过商业途径获得，但仍需要用于家畜研究，特别是牛研究的细胞系。在此，我们报告通过转染外源性人端粒酶逆转录酶（hTERT）建立了一种永生化牛肠上皮细胞（IBEC）系。从一头2日龄的杂种小牛中分离出原代牛肠上皮细胞（PBEC）。用hTERT转染PBEC以创建IBEC系。转染后的细胞对肠上皮标志物呈阳性，而对成纤维细胞标志物呈阴性。永生化后，IBEC的形态或生理特性没有显著差异。在跨上皮电阻（TEER测定）中，在脂多糖和丁酸钙处理下，PBEC和IBEC的表现与猪肠上皮细胞（IPEC-J2）相似。这种新开发的细胞系将成为体外牛和反刍动物肠道研究的重要且有用的工具。
