School of Veterinary Nursing and Technology, Faculty of Veterinary Science, Nippon Veterinary and Life Science University, Tokyo, 180-8602, Japan.
School of Veterinary, Faculty of Veterinary Science, Nippon Veterinary and Life Science University, Tokyo, 180-8602, Japan.
Sci Rep. 2021 Aug 10;11(1):16207. doi: 10.1038/s41598-021-95848-0.
Bovine-derived cultured cells, including Madin-Darby bovine kidney cells, are used worldwide; however, lipofection tend to result in low transfection efficiency, which has impeded the progress of veterinary research. We performed experiments to confirm the lipofection efficiency of bovine-derived cultured cells, to identify cells that suitable for lipofection. Several bovine tissues (endometrium, testis, ear tissue and foetal muscle) were collected, and primary cultured cells were prepared. Lipofection assay showed that only bovine endometrium (BE)-derived cells could be transfected efficiently (50‒70%). BE cells can be divided into at least two types of cell populations (BE-1 and BE-2). The BE-1 cells, which were suitable for lipofection, were obtained by passages at short intervals and were negative for cytokeratin- and positive for vimentin-expression; the BE-2 cells did not have these characteristics and were not suitable for lipofection. Furthermore, the BE-1 cells and artificially immortalised cells of BE-1, iBE-1 cells, were utilised in a reporter assay requiring the introduction of multiple DNAs. Endometrial tissues can be collected from living cows, and BE-1 cells can be obtained easily by controlling passaging timing. The production of BE-1 cells and sharing the methods required to prepare them will contribute to the development of veterinary research.
牛源培养细胞,包括 Madin-Darby 牛肾细胞,在全球范围内使用;然而,脂质体转染往往导致转染效率低,这阻碍了兽医研究的进展。我们进行了实验以确认牛源培养细胞的脂质体转染效率,以鉴定适合脂质体转染的细胞。收集了几种牛组织(子宫内膜、睾丸、耳组织和胎牛肌肉),并进行了原代培养细胞的制备。脂质体转染实验表明,只有牛子宫内膜(BE)来源的细胞可以高效转染(50-70%)。BE 细胞可分为至少两种细胞群(BE-1 和 BE-2)。通过短时间间隔传代获得适合脂质体转染的 BE-1 细胞,这些细胞对细胞角蛋白呈阴性,对波形蛋白呈阳性;而 BE-2 细胞则不具有这些特征,不适合脂质体转染。此外,我们还利用 BE-1 细胞和人工永生化的 BE-1 细胞 iBE-1 细胞进行了需要引入多种 DNA 的报告基因实验。可以从活体牛中收集子宫内膜组织,并通过控制传代时间来获得容易获得的 BE-1 细胞。BE-1 细胞的生产和分享制备它们所需的方法将有助于兽医研究的发展。