Cell culture and karyotype of Sakhalin sturgeon Acipenser mikadoi

A cell culture of rare and threatened species of Sakhalin sturgeon Acipenser mikadoi was established from a fragment of the pectoral fin and neighboring tissues. Initially, the culture consisted of different cell types including typical fibroblasts as well as cells of epithelial origin, myofibroblasts, etc. After approximately five passages, the culture largely consisted of cells with fibroblast morphology. Under normal culture conditions, these cells grew for more than one year at a constant rate and passed about 80 population doublings. In the absence of serum, cells entered the state of proliferative quiescence (G₀-state). When cultured without medium replacement for a long period of time, cells fused to form myofibers of about 1 cm in length. These myofibers could branch and acquired cross striation with time. About forty days after myofibers emerged, they degenerated, lost their shape, detached from the substrate, and finally died. The induction of adipogenic differentiation arrested cell proliferation and introduced lipophilic inclusions formed in a minor fraction of cells. The number of these inclusions was low, and cells with inclusions demonstrated various morphology distinct from typical adipocytes. The induction of osteogenic differentiation gave rise to cells that produce mineralized extracellular matrix and bone nodules. Chromosome analysis revealed a set of chromosomes typical for “high chromosome” sturgeon species. The variation in the chromosome number was very high (mean, 247 ± 33; modal value, 248). The analysis involving AT- and GC-specific fluorochromes has demonstrated that the telomeric and centromeric regions of all chromosomes are enriched in GC content. The distribution of AT- and GC-rich sequences along the chromosomes was heterogeneous. Long chromosomes were preferentially stained by the AT-specific dye, whereas small chromosomes demonstrated brighter fluorescence after 7-amino-actinomycin D staining; in particular, several small chromosomes fluoresced extremely brightly. This work is the first report of cell culture and karyotype analysis of Sakhalin sturgeon.