Characterization of Razi Bovine Kidney (RBK) cell line assensitive cell for BOHV-۱ virus

Background and Aim : Razi Bovine Kidney (RBK) cell line has been stabilized in Razi institute.RBK cell line is extremely sensitive to some viruses. This cell line was derived from primarybovine kidney cells by continuous passage. The purpose of this study is characterization of RBKcell line by identity tests (consist of karyology and molecular tests) and growth characteristics; toachieving an appropriate cell line for Bovine Herpesvirus ۱(BoHV-۱) virus.Methods : In order to preparing RBK cells, the standard methods were followed based on Freshney۲۰۱۶. Origin of RBK cell line was confirmed by PCR test. The RBK were incubated for differenttime periods such as ۲۴, ۴۸, ۷۲, ۹۶,۱۲۰, ۱۴۴, ۱۶۸ and ۱۹۲ hours, Then cell growth curve anddoubling time (DT) were determined. The cell DT was calculated using formula: T = t {lg۲ (lg Nt_ lg N۰)_۱} Cytogenetic analysis of RBK was carried out with standard procedure.Results : According to the single band in ۱۰۲ bp, the results of molecular identification revealedthat cell line origin was bovine. During DT the cell line exhibited an exponential growth pattern.Cells reached stationary growth phase after ۱۴۴ hours of exponential growth. Finally, the resultsof cell count and time of DT for RBK by using formula calculated. cytogenetic analysis revealedan aneuploid karyotype, that only ۴۲% of the RBK cells possessed the modal diploid chromosomenumber of ۲n=۴۴, which consisted ۴ pairs of metacentrics, ۲ pairs of subtelocentrics and ۱۶ pairsof telocentrics (۲n=۴m+۲st+۱۶t).Conclusion : Sensitivity of RBK cell line to different viruses and parasites was studied and variousresults have been reported. In this study, we found RBK cell line as the most effective forcultivation of BOHV-۱ virus, which is optimal for the propagation of BOHV-۱ virus. As well as,the BOHV-۱ virus titre reached ۸±۰.۵ CCID۵۰/mL on RBK cell line. Consider to DT and titrationtests, this research work proved that RBK has a recognizable and appropriate sensitivity to BOHV-۱ virus; In addition this cell line can be easily and rapidly proliferated.

کلیدواژه ها:

RBK ، DT ، cell line ، cytogenetic analysis ، aneuploid