Study on identification of Iranian BHK-21-C5 cell line by two steps PCR

Document Type: Original Articles


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

2 Department of Bio bank, Razi Vaccine & Serum Research Institute, Agricultural Research Education and Extension Organization (AREEO), Karaj, Iran

3 Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran.


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 that can be advantageous for functional use of animal cell lines. Therefore, efforts for prevention misidentified cells, cross-contamination to the other cell lines and mislabeling errors by using various procedures are continues and often leading to incorrect assessment. These contaminants can cause a massive financial disadvantage. One of the practical methods in this field is a molecular procedure that can show results more accurately. In this study, the BHK-21 (C5) characterized and we tried to determine the identity of BHK-21 (C5) as a continuous cell line in Iran by PCR molecular procedure and the cytochrome c oxidase I (CO1) gene was selected as a prevalent DNA fragment to authentication of BHK-21 (C5) cell line alongside six cell lines including CHO, LK, RBK, MRC5, VERO, GL with PCR and also to propagation of each cell line with its particular primer. After amplified, PCR products were analyzed by agarose gel electrophoresis to ensure their accuracy. The results of characterization indicated, cell viability was estimated to be about 92% and a uniform cell culture was obtained. The doubling time and µ ratio Equivalent respectively to 20.5 h and 0.03 were obtained. Sterility tests revealed that the cell seed was free of bacterial, mycoplasma and mycobacterial infections. The results of molecular identification show that the identification of this cell line was approved and can be used in researches, diagnosis studies and production and quality control of biological products.