Establishment and Characterization of Eye Muscle Cell Line from Snow Trout, Schizothorax richardsonii (Gray, 1832), a Vulnerable Coldwater Fish, for In Vitro Studies
Sanjeev Kumar, Pankaj Soni, Neha Singh, Ravindra Kumar, Shreya Srivastava, Akhilesh Kumar Mishra, Vijay Kumar Singh, Basdeo Kushwaha
Abstract
Schizothorax richardsonii, commonly called snow trout is a vulnerable coldwater fish distributed in India and adjoining countries, is an important resource as food, ornamental and game fishery. A cell line, designated as SREM-1, has been established from the eye muscle of S. richardsonii using explant method and cultured in Leibovitz’s L-15 medium. The growth studies at 20-32°C temperatures and 5-20% of fetal bovine serum (FBS) concentrations revealed that SREM-1 cells exhibited optimal growth at 28°C in L-15 medium containing 20% FBS. Cell type was confirmed primarily as epithelial in nature by immuno-phenotyping assay and was sub-cultured for >55 times since its development. The origin of the cell line was confirmed by sequencing of cytochrome oxidase c subunit I and 16S rRNA genes. SREM-1 cell line is free from mycoplasma contamination and has been cryopreserved at different passage levels with revival efficiency of 70-80% after 6 months. The SREM-1 cell line was successfully transfected and also employed for evaluating in vitro cytotoxicity against a heavy metal, mercuric chloride. The newly established cell line of this vulnerable species would be useful as a model for aquatic toxicological and transfection studies.