Laboratory of Molecular and Cellular Biology, Unit of Genetics, Faculty of Biological Sciences, University of Sciences and Technology "Houari Boumediene", Algiers, Algeria.
Laboratory of Production and Development of Viral Veterinary Vaccines, Pasteur Institute of Algeria, Algiers, Algeria.
In Vitro Cell Dev Biol Anim. 2021 Jan;57(1):76-85. doi: 10.1007/s11626-020-00520-y. Epub 2021 Jan 7.
Sheep primary epithelial cells are short-lived in cell culture systems. For long-term in vitro studies, primary cells need to be immortalized. This study aims to establish and characterize T immortalized sheep embryo kidney cells (TISEKC). In this study, we used fetal lamb kidneys to derive primary cultures of epithelial cells. We subsequently immortalized these cells using the large T SV40 antigen to generate crude TISEKC and isolate TISEKC clones. Among numerous clones of immortalized cells, the selected TISEKC-5 maintained active division and cell growth over 20 passages but lacked expression of the oncogenic large T SV40 antigen. Morphologically, TISEKC-5 maintained their epithelial aspect similar to the parental primary epithelial cells. However, their growth properties showed quite different patterns. Crude TISEKC, as well as the clones of TISEKC proliferated highly in culture compared to the parental primary cells. In the early passages, immortalized cells showed heterogeneous polyploidy but in the late passages the karyotype of immortalized cells became progressively stable, identical to that of the primary cells, because the TISEKC-5 cell line has lost the large SV40 T antigen expression, this cell line is a valuable tool for veterinary sciences and biotechnological productions.
绵羊原代上皮细胞在细胞培养系统中寿命较短。为了进行长期的体外研究，需要将原代细胞永生化。本研究旨在建立和鉴定 T 永生化绵羊胚胎肾细胞（TISEKC）。在本研究中，我们使用胎羊肾脏来获得原代上皮细胞培养物。随后，我们使用大 T SV40 抗原使这些细胞永生化，从而产生粗 TISEC 和分离 TISEC 克隆。在众多永生化细胞克隆中，选择的 TISEKC-5 在超过 20 个传代中保持活跃的分裂和细胞生长，但缺乏致癌的大 T SV40 抗原的表达。在形态上，TISEKC-5 保持其上皮特征类似于亲本原代上皮细胞。然而，它们的生长特性表现出截然不同的模式。与亲本原代细胞相比，粗 TISEKC 以及 TISEKC 克隆在培养中高度增殖。在早期传代中，永生化细胞表现出异倍体，但在后期传代中，永生化细胞的核型变得越来越稳定，与原代细胞相同，因为 TISEKC-5 细胞系已经失去了大 SV40 T 抗原的表达，该细胞系是兽医科学和生物技术生产的有价值的工具。
