Establishment, growth, cryopreservation and species of origin identification of three cell lines from white sturgeon, Acipenser transmontanus

Abstract

Three cell lines derived from fin (WSF), head soft tissue (WSHST) and body muscle (WSBM) were established from white sturgeon (Acipenser transmontanus). Characterization included determination of optimal growth kinetics, karyotyping, and mitochondrial ribosomal RNA (rRNA) genotyping. The primary cultures of these cells were generated by the explant technique using the L-15 medium supplemented with 20% fetal bovine serum and epidermal/fibroblast growth factors. The cells grew between 15-30 degrees C, but optimal growth occurred at 25 degrees C with a doubling time of 48 hours. The cell lines can be readily maintained in-vitro and have been subcultured over 35 times. Following cryopreservation in liquid nitrogen, thawed cells exhibit a viability of > 90% after a 16-month storage period. Chromosomal typing of these cell lines at their 17th passage revealed a chromosomal distribution of 242 to 278 with an apparent peak ranging from 250 to 260. Polymerase chain reaction amplification of mitochondrial 16S ribosomal RNA and sequence analysis indicated 100% identity of the sequences found in the cell lines with those found in the source animal, confirming that the cell lines were of A. transmontanus origin.