Development and characterization of a new muscle cell line developed from pearl spot, Etroplus suratensis (Bloch 1790)

Abstract

Cell lines originating from diverse fish species inhabiting freshwater, brackish water, and marine environments have emerged as an innovative in vitro research tool in bioscience. A new cell line was developed from the muscle tissue of Etroplus suratensis (Pearl spot). The primary muscle cell culture system was successfully obtained from the dorsal muscle tissue was used for explant preparation of Etroplus suratensis and subsequently subcultured up to 15 passages, resulting in the development of a novel muscle cell line termed ESM (Etroplus suratensis muscle). Initially, the ESM cells were cultured in Leibovitz's 15 media (L-15) supplemented with 20 % fetal bovine serum (FBS) at a temperature of 28 °C. Muscle cells were then optimized for growth in culture media by reducing the FBS concentration to 15 % FBS and 5 % FBS. Additionally, basic fibroblast growth factor (bFGF), at a concentration of 10 ng/mL, was added to the cell culture flask so that the proliferation capacity of the ESM cell line could be increased and also reduce the time interval of subculture at different passages. Immunocytochemistry was carried out to understand the expression of myogenic regulatory factors MyoD and Pax7 in the developed cell line. The transfection efficiency of the ESM cells was 9 % using the pMaxGFP vector. The cytotoxicity assay of three heavy metal salts (Cd, Cu, Pb) was assessed in ESM cells using XTT and MTT assays. The COI gene was used to authenticate the species of origin of the developed ESM cell line. The ESM cell line was cryopreserved in liquid nitrogen (−196 °C). This is the first report of a muscle cell line derived from the fish species Etroplus suratensis.