Isolation of cells from atlantic sturgeon acipenser oxyrinchus oxyrinchus and optimization of culture conditions

Abstract

We describe a method for fast and easy isolation of cells via trypsin digestion from larvae of Atlantic sturgeon Acipenser oxyrinchus oxyrinchus resulting in a stable, well-proliferating cell culture. The culture conditions for these cells were optimized with the aim of supporting the production of high amounts of biomass. To enhance cell growth and cell density, 4 different cultivation temperatures as well as commercially available carp serum (CS) and fetal calf serum (FCS) at different concentrations were tested and evaluated. Cell growth was measured via an impedancebased online cell-monitoring system (xCELLigence). These results showed the best cultivation temperature to be at 25°C and a media composition of Dulbecco’s modified Eagle’s medium (DMEM) supplemented with either 10 or 20% FCS or 5% CS. The cells were stable in the process of longterm cultivation over 33 passages and could be cryo-preserved. Immunocytochemical analysis revealed that the cells expressed proteins of different blastodermic layers. Ectodermic glia fibrilliary acid protein, vigilin (mRNA transport protein), and pan cytokeratin were abundant. This fastgrowing cell culture provides an important tool for research on Atlantic sturgeon populations.