Establishment and Characterization of Novel Channa striata Muscle Cell Line for Scaffold-Based Micro-tissue Production

Abstract

Channa striata is a hardy fish that can thrive in various aquatic conditions; it is a good source of protein and has high economic value. This study was conducted to establish and characterize the muscle cell line of C. striata and use it to develop micro-tissue using seaweed biofilm. Explant culture was used to create a continuous muscle cell line from C. striata, which was then sub-cultured 143 times in Leibovitz L-15 medium with 10% FBS at 28 °C. They were properly cryopreserved, and after storage, 88–92% of the cells were recovered. The C. striata muscle cell line (CSM) was authenticated by examining the mitochondrial 16S rRNA gene using polymerase chain reaction. The immunophenotyping results revealed that the CSM cells were of myoblast origin, as evidenced by desmin and myosin cell markers. CSM cells showing a myogenic-like phenotype were also demonstrated through lipid droplets, which were confirmed by Nile Red staining. This cell line was devoid of mycoplasma infection. Inducing the foreign gene plasmid pEGFP-N1 resulted in a 16% transfection efficiency. Sterilized biocompatible seaweed biofilm was employed as a scaffold to grow micro-tissue using the developed CSM cell line. After 20 days of growth, this cell line produced micro-tissue on the seaweed biofilm. Seaweed biofilm is a one-of-a-kind biomaterial that has a wide range of biomedical applications, is ecologically friendly, biocompatible, and helps to improve cellular aquaculture and sustainable development.