Development and partial characterization of primary cell culture from liver tissue of Pangasianodon hypophthalmus.

Abstract

The present study was undertaken to optimize the conditions for development of primary hepatocyte cultures from Pangasionodon hypophthalmus. Liver tissue explants were used as cell seeding source and a protocol for liver cell culture was optimized considering four parameters i.e. incubation temperature, culture media, pH of the media and serum concentration. Among growth media, L-15, Hepatocyte Basal Medium, DMEM and RPMI-1640 with varying concentration of Foetal Bovine Serum (FBS) were tested for their ability to support the growth of cells. Later, the effect on cell confluence at different pH and at different incubation temperature was monitored. The cells were primarily characterized by Best’s carmine staining solution, which was later partially characterized by amplification of mt 16S rRNA gene fragment of 546 bp size that confirmed its origin from the liver explants of P. hypophthalmus at 20th passage. This study proved that the L-15 medium with pH 7.6 and 20% of serum concentration at incubation temperature of 28 0C yielded the best result in terms of attachment and proliferation of cells which can be used to develop the liver cell line or as a screening tool for further cytological studies.