Establishment of a Novel Muscle Cell Line From Wallago attu for In Vitro Study of Pesticide Toxicity

Abstract

species. Objectives: The present study aims to establish and characterize a novel fish cell line from muscle tissue of Wallago attu catfish, while evaluating its potential as an in vitro system for toxicity testing. Materials and Methods: Explant culturing method was used for the establishment of a cell line in Leibovitz-15 medium with 20% fetal bovine serum (FBS). The resulting cell line was characterized for optimal growth parameters, authenticity, stability, cellular morphology and revival efficiency. The conformity of the cell line for applied studies was established by using gene transfection and cytotoxicity studies against the organophosphate pesticides chlorpyrifos and malathion. Basic fibroblast growth factor was supplemented at 10 ng/ mL concentration. The origin of the cell line was authenticated using homology analysis of amplified gene sequences of 16S rRNA and mitochondrial Cytochrome Oxidase Subunit I (COI) gene. Cytotoxicity assessment of the pesticides was assayed by using methylthiazol tetrazolium assay (MTT) and neutral red uptake. Results: The resulting muscle cell line was designated as Wallago attu Muscle (WAM) and was maintained for over 50 passages at optimal growth temperature of 28°C. Chromosomal analysis confirmed the stability of the cell line. The cells exhibited fibroblastic morphology with good transfection and revival efficiencies. Conclusions: The established muscle cell line designated as WAM presented the stability while possessing good transfection and revival efficiency. These characteristics confirmed through cytotoxicity assessment of the pesticides using methylthiazol tetrazolium assay and neutral red uptake support the availability of this cell line as an in vitro system for toxicity evaluation of aquatic pollutants.