Isolation of cells from Atlantic sturgeon Acipenser oxyrinchus oxyrinchus and optimization of culture conditions

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 impedance-based 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 long-term 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 fast-growing cell culture provides an important tool for research on Atlantic sturgeon populations.