Establishment and Molecular Characterization of a Liver-derived Cell line from Schizothorax esocinus (Chirruh) for Cold Water Fish Research

Abstract

Schizothorax esocinus, commonly known as Chirruh, is an important indigenous coldwater fish of the Kashmir. The new cell line was developed from the liver tissue of Schizothorax esocinus, and was characterized by molecular technique. The developed cell line was maintained and passaged 14 times in Leibovitz’s L-15 medium. The cell line was standardized using L-15 media by testing at various FBS (fetal bovine serum) concentrations and temperature ranges. Cell numbers increased more rapidly in 20% FBS, compared to other concentrations ranging from 5% to 25%. The tissue got fully adhered after 72hrs when seeded at 24°C as compared to other temperatures ranging from 8 to 28°C. The tissue preparation approach was more reliable than the enzymatic dissociation approach. Cell colonies of various sizes were formed after 1 week and 60-70% confluent cell monolayer were achieved after 20 days. The cell morphology was found to be of epithelial-like cells. The origin of the cell line was confirmed at passage 12 by the amplification of 629 bp of cytochrome oxidase subunit I (COI) similar to the original liver issue. The current cell line developed from liver tissue of S. esocinus has tremendous potential to enhance the generation of cold-water fish cell lines could and potentially be used as a important approach for conservation genetics and biotechnological applications.