IMMORTALIZATION OF SV40-TRANSFORMED HUMAN CELLS

SV40 转化人类细胞的永生化

• 批准号: 3115384
• 负责人: HARVEY L OZER
• 金额: $ 27.8万
• 依托单位: UNIV OF MED/DENT OF NJ-NJ MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 1988
• 资助国家: 美国
• 起止时间: 1988-12-01 至 1992-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3115384
• 关键词: aging; cell age; cell bank /registry; chromosome aberrations; clone cells; diploidy; fibroblasts; gene dosage; gene expression; genetic manipulation; genetic recombination; human tissue; karyotype; mutagens; nucleic acid probes; oncogenic virus; radiotracer; simian virus 40; temperature sensitive mutant

Human diploid cells derived from normal tissue undergo a characteristic pattern of growth in cell culture, ending with a non-replicative phase and, finally, cell death. Models of cellular aging have been developed which suggest analogy between this pattern and behavior of the organism in vivo; hence, the term "senescence". Human diploid fibroblasts (HF) which had been infected in vitro by the oncogenic DNA virus SV40 have been found to overcome this phenomenon. We have found that a clonal isolate of a HF transformed by an origin-defective mutant of SV40 (SVori-) with an intact early region permits isolation of immortalized cells at an unusually high frequency. Analysis of several such immortalized HF from individual cloned transformants will be performed to identify possible common genetic alterations. Further, we propose to study a series of independent HF transformed by origin-defective mutants of SV40 with a temperature-sensitive T antigen (ts A mutants) to assess possible bases for the role of SV40 in the initiation and maintenance of a continuous cell strain.

来自正常组织的人二倍体细胞经历一个特征性的 细胞培养中的生长模式，以非复制期结束， 最后是细胞死亡。 已经开发了细胞老化模型， 提出这种模式和生物体在体内的行为之间的相似性; 因此，术语“衰老”。 人二倍体成纤维细胞（HF）， 在体外被致癌DNA病毒SV 40感染， 克服这种现象。 我们发现HF的克隆分离物 由SV 40的来源缺陷突变体（SVori-）转化， 早期区域允许以异常高的浓度分离永生化细胞， 频率. 几个这样的永生化的HF从个体克隆的分析 将进行转化以鉴定可能的共同遗传 改变。 此外，我们建议研究一系列独立的HF 由SV 40的来源缺陷突变体转化， 温度敏感性T抗原（ts A突变体），以评估 SV 40在启动和维持连续细胞中的作用 株