ADAPTATION AND ANALYSIS OF PORCINE CIRCOVIRUS 2 IN PORCINE KIDNEY (PK-15) CELLS CULTURED WITH LOW SERUM SUPPLEMENTATION

A. Kalaimathi; S. Parthiban; P. Raja; J. Vijay Anand; M. Parthiban

doi:10.56093/ijvasr.v54i4.169945

Authors

A. Kalaimathi M.Sc Scholar, Department of Biotechnology, School of Biological Sciences, CMS College of Science and Commerce, Coimbatore
S. Parthiban Assistant Professor, Department of Animal Biotechnology, Faculty of Basic Sciences, TANUVAS, Madras Veterinary College, Chennai - 600 007
P. Raja Assistant Professor, Department of Animal Biotechnology, Faculty of Basic Sciences, TANUVAS, Madras Veterinary College, Chennai - 600 007
J. Vijay Anand Assistant Professor, Department of Animal Biotechnology, Faculty of Basic Sciences, TANUVAS, Madras Veterinary College, Chennai - 600 007
M. Parthiban Professor and Head, Department of Animal Biotechnology, Faculty of Basic Sciences, TANUVAS, Madras Veterinary College, Chennai - 600 007

https://doi.org/10.56093/ijvasr.v54i4.169945

Keywords:

PK-15 cell, Serum, deprivation PCV2, adaptation, genetic analysis

Abstract

This study aimed to cultivate PK-15 cells under serum-deprived conditions and subsequently adapt a field-isolated PCV2b genotype in these adapted cells to investigate both cellular and viral changes. PK-15 cells were grown in three different serum-supplemented media: 0%, 2%, and 10%. The cells adapted to these varying serum conditions were then infected with genotyped field PCV2b and propagated for up to three blind passages. Haematoxylin and eosin (H&E) staining of PK-15 cells grown in 0% and 2% serum-supplemented media, with and without PCV2b infection, revealed cytoplasmic granularity compared to cells grown in 10% serum. MTT assay results indicated that PCV2b infection had minimal impact on cell viability, suggesting efficient viral propagation in serum-deprived conditions with minimal cytopathic effects. PCR analysis of cell lysates from all three 0%, 2%, and 10% serum conditions confirmed the presence of the PCV2b genome, demonstrating its adaptability grown to PK-15 cells under serum-deprived conditions. Molecular analysis of the major epidemiological marker gene, ORF2, from PCV2b grown in both 0% and 10% serum conditions showed identical nucleotide (702 bp) and amino acid (234 aa) compositions. This study highlights the feasibility of growing PK-15 cells under serum-deprived conditions and the adaptability of PCV2b in these cells.

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