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ISSN : 2287-7991(Print)
ISSN : 2287-8009(Online)
Journal of the Preventive Veterinary Medicine Vol.38 No.1 pp.15-25

Immortalization of bovine primary epithelial cells to isolate foot-and-mouth disease virus

Kyu-Kye Hwang, Kyong-Leek Jeon
College of Veterinary Medicine, Jeju National University, Jeju 690-756, Republic of Korea
Received 02 February 2014, Revised 08 March 2014, Accepted 24 March, 2014


A cell line of bovine origin was immortalized to isolate foot-and-mouth disease virus (FMDV). The immortalizationwas performed by infection of bovine primary epithelial cells with a recombinant retrovirus that overexpressed the humantelomerase (hTERT), after primary culture of fetal bovine kidney tissue and removal of fibroblasts. After cloning the immortalizedcell line into single cells, the cloned cell lines were named JNUBK-1, JNUBK-2, JNUBK-3 and JNUBK-4, according totheir characteristics. To confirm the epithelial phenotype of the cell lines JNUBK-3 and JNUBK-4, which showed stableproliferation capability over 35 generations after immortalization, the expression of cytokeratin and fibronectin was measured.Finally, the FMDV titer in the JNUBK-3 and JNUBK-4 cell lines was measured and was 800∼2,000 times higher than thatof the currently used cell line IRBS-2. In conclusion, more sensitive isolation and production of FMDV became possiblethrough the use of the immortalized JNUBK-3 and JNUBK-4 cell lines.