Development and Cytogenetic Characterization of a Continuous Bovine Kidney Cell Line (IRKHBK) and Evaluation its Susceptibility to some Viruses

در اين تحقيق يك تيره ياخته اي مداوم از سلول اوليه كليه گوساله براي اولين بار در ايران توليد شد. منشا اين سلول يك گوساله نر 2 روزه طبيعي نژاد هولشتاين بود. سلول اوليه كليه گوساله بعد از رشد كامل بطور مداوم پاساژ داده شد. با بهره گيري از آناليزهاي سيتوژنتيك خصوصيات ژنتيكي سلول تعيين و عدم تومورزايي سلول ثابت شد. در پاساژ سي ام سلول، افزايش تكثير سلولي مشاهده شد. آناليز كروموزم ها اولين چسبيدگي كروموزمي را نشان داد. در كاريوتايپ و در مقايسه با كروموزوم هاي سلول طبيعي گوساله (2n= 30) تعداد كروموزوم هاي سلول به 59 عدد (n = 59) كاهش يافت. از هفتادمين پاساژ توانايي تكثير سلول ها نامحدود شد و از پاساژ نود به بعد به عنوان تيره ياخته اي تعيين شد. اين تيره ياخته اي مداوم به نام Iran Razi Khedmati Bovine Kidney (IRKHBK) نامگذاري و در بانك سلولي ايران (NCBI) وابسته به انستيتوپاستور ايران ثبت شد حساسيت تيره سلولي IRKHBK براي جداسازي و تكثير bovine herpesvirus-1 (BHV-1) bovine virus و diarrhea-mucosal disease (BVD-MD) ارزيابي گرديد. نتايج نشان داد كه حساسيت اين سلول در مقايسه با سلول اوليه BK نسبت به اين ويروس ها بيشتر مي باشد. مطابق نتايج اين بررسي، سلول IRKHBK براي روش هاي معمول ويروس شناسي بعنوان جايگزين سلول اوليه BK توصيه مي شود.

In this syudy a continuous bovine kidney cell line derived from a primary bovine kidney cells was established for the first time in Iran. The cells were originating from two-day-old normal male calf of Holstein breed. The cell cultures were continuously passaged following complete proliferation of primary cells. The specific properties or characteristics of the cell were defined using cytogenetic and tumorigenicity analysis. An increasing in cell proliferation was observed at 30th passage. Subsequently chromosomes analysis was shown the first chromosomal adhesion. In karyotyping a decrease in number of the cell¢s chromosomes (n=59) was detected compared to the normal bovine cells chromosome count (2n=30). The cell obtained unlimited proliferation capacity from 70th passage and was identified as infinite cells at passage 90th. The continuous cell line named Iran Razi Khedmati Bovine Kidney (IRKHBK) and was deposited in National Cell Bank of Iran (NCBI), Pasteur Institute. Susceptibility of the IRKHBK cell line for isolation and replication processes of bovine herpesvirus-1 (BHV-1) and bovine virus diarrhea-mucosal disease (BVD-MD) were also evaluated. The results showed that this cell is more susceptible to the viruses compared to primary bovine kidney cells. According to our results, IRKHBK cell is recommended for routine assays of viruses as a substitution for primary bovine kidney cells.