An optimization of supplements and physical factors for growth of hemocytes culture from Penaeus vannamei (White shrimp) in selective medium.

BACKGROUND

Standardization of cell culture medium plays a vital role in the development of primary or continuous cell line. Apart from the basal media, supplements in the medium and various physical factors promote the cell growth. With this context, the study was carried out to optimize the culture medium using various supplements and physical factors for the growth of hemocytes culture from Penaeus vannamei.

METHODS

Various concentrations of Fetal Bovine Serum (FBS; 1-25%), Shrimp Muscle Extract (SME; 1-25%) and basic Fibroblast Growth Factor (bFGF; 0.5-5 ng mL ) were attempted to optimize the cell culture media for the development of primary hemocytes culture of P. vannamei. Various pH, temperature and osmolality was also screened to optimize the medium.

RESULTS

15% FBS was ideal for the healthy morphology of cells with rapid replication. SME supplementation at 5-20% supported the cell growth for 24 h but only 30% of cell viability was observed after 48 h. bFGF (0.5-5 ng mL) enhanced cell growth in the medium with 15% FBS; The ideal pH level was examined by preparing the HBSCM-5 medium at pH between 6.8-8.0. Osmolality of 730 ± 20, pH of 7.2 and temperature of 28 °C resulted in the healthy cells with good morphology. NSW supplement supported the cell growth at low concentrations of salt; however, more than 2% salt concentrations cells did not form fibroblast-like morphology and instead a crystal-like morphology was observed.

CONCLUSION

The hemocytes culture were optimized for use as an in vitro cell culture system by testing cell growth on HBSCM-5 medium with various supplements, growth factors and physical parameters.