Rao J.P.,Harrison Center for Advanced Research in Biotechnology |
Rao S.K.R.S.,Acharya Nagarjuna University
Research Journal of Biotechnology | Year: 2011
A primary cell culture system was developed for the cells of hepatopancreatic tissue of Penaeus vannamei post larvae. Minced tissue fragments of hepatopancreas were disaggregated with 0.1% trypsin. The cells were seeded at different densities in three different media viz. minimum essential medium (MEM), Hanks balanced salt solution (HBSS) and L15 medium. Epitheliod like cells were developed in all media. The cultured conditions like time of disaggregation, seeding density, temperature, CO 2 concentrations were optimized. The primary cell cultures were also established by supplementing all the three media with different biological fluid extracts like Fetal bovine serum (FBS), Prawn muscle extract (PME) and Fish fry extract (FFE).Maximum number of viable cells were obtained with 30 minutes disaggregation and when cells were seeded at a density of 10 6 cells/ml, primary cell cultures were best established in L-15 medium among the three media used with different biological fluids like FBS (20%), prawn muscle extract (PME) and fish fry extract (25%). They were survived for a period of 50 days. Cell cultures were best survived with 5% CO 2 and at 30°C temperature.