Rolland Morgane, Chauvineau Cécile, Valas Stephen, Mamoun Robert Z, Perrin Gérard
AFSSA-Niort, Laboratoire d'Etudes et de Recherches Caprines, 79012 Niort, France.
J Virol Methods. 2004 Jun 15;118(2):123-30. doi: 10.1016/j.jviromet.2004.02.001.
Primary goat synovial membrane (GSM) cells are widely used to study small ruminant lentiviruses (SRLV), i.e. maedi visna virus (MVV) and caprine arthritis-encephalitis virus (CAEV), but their limited life-span of 15-20 passages in vitro is problematic. Here, we report that ectopic expression of the catalytic subunit of human telomerase (hTERT) was sufficient to immortalize primary GSM cells. Cultures of hTERT-transfected GSM cells have been passaged for 2 years without showing any phenotypic difference from the original primary GSM cells. The hTERT-transfected cells continued to grow beyond a population doubling number of 250, while no net telomere lengthening was observed for these cells. Moreover, the immortalized GSM cells were susceptible to infection by both CAEV and MVV and were able to propagate theses viruses. Such cell line provides a useful source of standard and robust cells for both research and veterinary purposes.
原代山羊滑膜(GSM)细胞被广泛用于研究小反刍兽慢病毒(SRLV),即梅迪-维斯纳病毒(MVV)和山羊关节炎-脑炎病毒(CAEV),但其在体外有限的传代寿命(15-20代)存在问题。在此,我们报告人端粒酶催化亚基(hTERT)的异位表达足以使原代GSM细胞永生化。hTERT转染的GSM细胞培养物已传代2年,与原始原代GSM细胞相比未显示任何表型差异。hTERT转染的细胞在群体倍增数超过250后仍继续生长,而这些细胞未观察到端粒净延长。此外,永生化的GSM细胞对CAEV和MVV感染均敏感,并能够传播这些病毒。这种细胞系为研究和兽医目的提供了有用的标准且稳定的细胞来源。