بررسی سنجش فعالیت آنزیم تلومراز در سلول ‭MRC5 ‬مورد استفاده در موسسه رازی و مقیسه آن با سلول ‭Hella‬

MRC-5 is a diploid cell with limited life span which used as a cell substrate in viral culture e.g. Measles and Polio virus. Telomerase and telomeres have recently studied as a biological clock for control of cell proliferation and aging. The high level of telomerase expression in about 90% of human tumors is detected and suggested as a tumor marker. However, few studies have addressed telomerase activity in diploid cell. In this study telomerase activity during cell senescence of human MRC-5 cells as cellular substrate for Measles virus propagation was investigated. Telomerase activity assayed in different passage number from 15 to 38 of this cell with and without viral transfection. The activity determined with a modified TRAP-PCR assay and the result evaluated with both PAGE gel and ELISA simultaneously. The gels stained either with ethidium bromide or SYBR Green. The SYBR Green staining was better than ethidium bromide because of its higher sensitivity. The best gel was 20% PAGE for separation of PCR products in TRAP-PCR technique. In general, trace telomerase activity detected in these cells but this trace telomerase activity of MRC-5 decreased with increasing of passage number. The results also showed that infected MRC-5 cells had relatively higher activity. Karyotype of cells showed that most of cells were normal. Titeration of AIK-C Measles was decreased with senescence increase of age of MRC-5. Key words: Cell culture, Telomerase, Telomere, Measles virus aging, TRAP-PCR, TRAP-ELISA 78

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سلول های هلا ، دیپلویید ، سرخک ، ویروس ، تومورهای انسانی ، آنزیم های تلومراز