مقالات پذیرفته شده در هفتمین کنگره بین المللی زیست پزشکی
Characterization of RBK cell line as a sensitive cell to bovine Herpesvirus-1 (BoHV-1)
Characterization of RBK cell line as a sensitive cell to bovine Herpesvirus-1 (BoHV-1)
Masoumeh Maghami,1,*Mohsen Lotfi,2Fattah Sotoodehnejadnematalahi,3Ashraf Mohammadi,4
1. Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran 2. Department of Quality Control, Razi Vaccine and Serum Research Institute, Agricultural Research, Education, and Extension Organization (AREEO), Karaj, Iran 3. Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran 4. Department of Human Viral Vaccines, Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran
Introduction: Viruses are obligate parasites and are completely dependent on host cells for survival and replication. RBK cell line developed and introduced by Razi Vaccine and Serum Research Institute, has been successfully established as a continuous cell line over successive passages.
Methods: RBK cells demonstrate marked sensitivity to certain viruses. In this experimental study, RBK cell line has shown significant sensitivity to BoHV-1 virus. At present research, the characteristics of RBK cell line were performed by molecular and karyotype methods, as well as growth characteristics. Cloning of the RBK cell line was performed using limited dilution method in parallel with analytics for characterization of each cell clone quantitatively and qualitatively. Then four cell clones were compared based on their sensitivity to the BoHV-1 virus.
Results: in this study, the RBK-D5 clone was selected as the most appropriate cell line for further study and was subjected to tests for identity, chromosomal analysis and doubling time. In the end, RBKD5 in terms of species origin and contamination-free was confirmed by PCR method, as well as RFLP-PCR and Real-time PCR. We observed that the cell line directed towards karyotype diversity, because of aneuploidy. Aneuploidy itself can be responsible for the procreation of chromosomal instability. Karyotype diversity represent chromosomal changes in the continuous cell line that carries the characteristic of immortalized cell line.
Conclusion: In general, one of the key factors in virus isolation is multiplication speed in cell culture and high production efficiency. Therefore, it's absolutely necessary to choosing a suitable cell line with high sensitivity to certain virus. At present study, our results revealed that RBK-D5 cell line contain high growth rate, stability and surprisingly sensitive to BOHV-1 virus at different passages, as well as our results confirmed the absence of contamination to other cells and unwanted factors.
In total, the conducted investigations confirm the capability of the studied cell. As a conclusion the RBK cell line can be widely used in research, detection, virus propagation and virus quality control or titration.