Institute of Cell Biology and Neurobiology, National Research Council, Rome, 00143, Italy.
Institute of Medical Pathology, Università Cattolica, Rome, 00168, Italy.
Endocrine. 2018 Jan;59(1):102-108. doi: 10.1007/s12020-017-1305-6. Epub 2017 Apr 26.
The aim of this study was to generate immortalized human anterior pituitary adenoma cells. Reliable cell models for the study of human pituitary adenomas are as yet lacking and studies performed so far used repeated passaging of freshly excised adenomas, with the attendant limitations due to limited survival in culture, early senescence, and poor reproducibility.
METHODS & RESULTS: We devised a technique based upon repeated co-transfections of two retroviral vectors, one carrying the catalytic subunit of human telomerase, hTERT, the other SV40 large T antigen. This approach extended the lifespan of cells derived from a human growth hormone-secreting adenoma up to 18 months while retaining morphology of primary cells, growth hormone synthesis and growth hormone secretion.
Our attempt represents the first demonstration of successful lifespan extension of human growth hormone-secreting pituitary adenoma cells via co-transfection of hTERT and SV40T and paves the way to future attempts to obtain stable cell lines.
本研究旨在生成永生化的人垂体腺瘤细胞。目前仍然缺乏可靠的垂体腺瘤细胞模型，到目前为止进行的研究使用了反复传代的新鲜切除的腺瘤，由于在培养中生存能力有限、早期衰老和重现性差，因此存在一定的局限性。
我们设计了一种基于两次逆转录病毒载体共转染的技术，其中一个载体携带人端粒酶催化亚单位 hTERT，另一个载体携带 SV40 大 T 抗原。这种方法将来源于生长激素分泌型垂体腺瘤的细胞寿命延长至 18 个月，同时保留了原代细胞的形态、生长激素的合成和生长激素的分泌。
我们的尝试首次成功地通过 hTERT 和 SV40T 以及 p 的共转染延长了人生长激素分泌型垂体腺瘤细胞的寿命，为未来获得稳定细胞系的尝试铺平了道路。
