Identification of Iranian BHK-۲۱-C۵ Cell Line by Two Steps Polymerase Chain Reaction

سال انتشار: 1400
نوع سند: مقاله ژورنالی
زبان: انگلیسی
مشاهده: 69

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شناسه ملی سند علمی:

JR_ARCHRAZI-76-2_002

تاریخ نمایه سازی: 6 دی 1402

چکیده مقاله:

Authentication of animal cell lines in cell banks is one of the most important programs regulated during cell culture and storage. This operation provides a thorough and beneficial document which can be advantageous for the functional use of animal cell lines. Therefore, various procedures are used to prevent misidentified cells, cross-contamination to other cell lines, and mislabeling errors leading to incorrect assessment. These contaminants can result in major financial disadvantages. One of the practical methods in this field is a molecular procedure which can demonstrate more accurate results. In the present study, the BHK-۲۱ (C۵) was characterized, and it was tried to determine the identity of BHK-۲۱ (C۵) as a continuous cell line by Polymerase chain reaction (PCR) molecular procedure in Iran. The cytochrome c oxidase I (CO۱) gene was selected as a prevalent DNA fragment for the authentication of the BHK-۲۱ (C۵) cell line, along with six cell lines, including Chinese hamster ovary,  Lamb kidney, Razi Bovine Kidney, Medical Research Council cell strain ۵, Monkey Green Kidney, and Goat Lymphocyte. After amplification, PCR products were analyzed by agarose gel electrophoresis to ensure their accuracy. The results of characterization were indicated, cell viability was estimated to be about ۹۲%, and a uniform cell culture was obtained. The doubling time and µ ratio equivalent were obtained at ۲۰.۵ h and ۰.۰۳, respectively. Sterility tests revealed that the cell seed was free of bacterial, mycoplasma, and mycobacterial infections. The results of molecular identification revealed that the identification of this cell line was approved and can be used in studies, diagnosis, production, and quality control of biological products.

کلیدواژه ها:

Authentication ، BHK-۲۱-C۵ cell line ، PCR ، cytochrome c oxidase I (CO۱) gene

نویسندگان

F Ziyaeifar

Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran

S Soleimani

Department of Biobank, Razi Vaccine and Serum Research Institute, Agricultural Research Education and Extension Organization (AREEO), Karaj, Iran

M Lotfi

Department of Viral Vaccine Quality Control, Razi Vaccine and Serum Research Institute, Agricultural Research Education and Extension Organization (AREEO), Karaj, Iran

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