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Characterization of Razi Bovine Kidney (RBK) Cell Line as a Sensitive Cell to Bovine Herpesvirus-1 (BoHV-1) | ||
| Archives of Razi Institute | ||
| مقاله 14، دوره 78، شماره 5، آذر و دی 2023، صفحه 1528-1541 اصل مقاله (1.58 M) | ||
| نوع مقاله: Original Articles | ||
| شناسه دیجیتال (DOI): 10.32592/ARI.2023.78.5.1528 | ||
| نویسندگان | ||
| Masoumeh Maghami1؛ Mohsen lotfi* 2؛ Fattah Sotoodehnejadnematalahi1؛ Ashraf Mohammadi3 | ||
| 1Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran | ||
| 2Department of Quality Control, Razi Vaccine and Serum Research Institute, Agricultural Research, Education, and Extension Organization (AREEO), Karaj, Iran. | ||
| 3Department of Human Viral Vaccines, Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran | ||
| چکیده | ||
| 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.Viruses are obligate parasites that completely rely on host cells for survival and replication. Razi Bovine Kidney (RBK) cell line was introduced and developed by the Razi Vaccine and Serum Research Institute. It has been successfully established as a continuous cell line over successive passages. As demonstrated in this experimental study, the RBK cells have shown suitable sensitivity to certain viruses, including Bovine Herpesvirus-1 (BoHV-1) virus. In the present research, the RBK cell line characteristics were analyzed using molecular and karyotype methods and growth specifications. Cloning of the RBK cell line was performed using the limited dilution method, and each cell clone was quantitatively and qualitatively characterized. Four cell clones were compared based on their sensitivity to the BoHV-1 virus. Then, the RBK-D5 clone was selected as the most appropriate cell line for further studies. The RBK-D5 was subjected to tests for identity, chromosomal analysis, and doubling time. In the end, the origin of this cell line was confirmed by the PCR method. It was observed that the cell line exhibited karyotype diversity due to aneuploidy, which can be responsible for the procreation of chromosomal instability. This diversity represents chromosomal changes in the continuous cell line that carries the characteristic of an immortalized cell line. The RBK-D5 was found to be more sensitive to the BOHV-1 virus. Surprisingly, its titer was evaluated at 108.5 CCID50/ml. The obtained results suggested that the RBK cell line is suitable for the BoHV-1 virus and can be useful for virus detection, propagation, and quality control or viral titration. | ||
| کلیدواژهها | ||
| BoHV-1 virus؛ Karyotyping؛ RBK cell line؛ RBK-D5 | ||
| عنوان مقاله [English] | ||
| تعیین ویژگی های رده سلولی RBK به عنوان یک سلول حساس به ویروس هرپس گاوی تیپ یک (BoHV-1) | ||
| چکیده [English] | ||
| ویروس ها انگل های اجباری هستند که برای بقا و تکثیر کاملاً به سلول های میزبان متکی هستند. رده سلولی RBK که توسط موسسه تحقیقات واکسن و سرم سازی رازی تهیه و معرفی شده است، با موفقیت به عنوان یک رده سلولی پیوسته متعاقب پاساژ های متوالی ایجاد شده است. مطالعات تجربی نشان داده است، سلولهای RBK حساسیت مناسبی نسبت به ویروسهای خاص، از جمله ویروس BoHV-1 نشان می دهند. در تحقیق حاضر، ویژگیهای رده سلولی RBK با استفاده از روشهای مولکولی و کاریوتیپ و همچنین شاخصه های رشد مورد تجزیه و تحلیل قرار گرفت. شبیه سازی رده سلولی RBK با استفاده از روش رقت گیری محدود انجام شد و هر کلون سلولی از نظر کمی و کیفی ارزیابی گردید. چهار کلون سلولی بر اساس حساسیت آنها به ویروس BoHV-1 مقایسه شد. سپس کلون RBK-D5 به عنوان مناسب ترین رده سلولی برای مطالعه بیشتر انتخاب شد. RBK-D5 تحت آزمایش های هویت، تجزیه و تحلیل کروموزومی و زمان دو برابر شدن قرار گرفت. در نهایت منشاء این رده سلولی با روش PCR تایید شد. مشاهده شد که رده سلولی مذکور به دلیل آنئوپلوئیدی، تنوع کاریوتیپ را نشان میدهد که میتواند مسئول تولید بیثباتی کروموزومی باشد. این تنوع نشان دهنده تغییرات کروموزومی در رده سلولی پیوسته است که ویژگی یک رده سلولی نامیراست. RBK-D5 به ویروس BOHV-1 حساس تر است. با کمال تعجب، تیتر آن CCID50/ml 108.5 ارزیابی شد. این نتایج نشان می دهد که رده سلولی RBK برای ویروس BoHV-1 مناسب است و می تواند برای تشخیص، تکثیر و یا تیتراسیون ویروس مفید باشد. | ||
| کلیدواژهها [English] | ||
| سلول لاین RBK, ویروسBoHV-1, کاریوتایپینگ, RBK-D5 | ||
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| مراجع | ||
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