Studies on cryopreservation of luc gene transfected bluegill sunfish fibroblast cell line.
The effect of cryopreservation on the survival of luc gene transfected bluegill sunfish fibroblast (BF-2) cells was investigated. Propane-1,2-diol was found to be the least toxic cryoprotectant when compared with DMSO and methanol. Both propane-1,2-diol and DMSO are effective in protecting cells from freezing damage. Whilst there were no significant differences in cell survival between cryoprotectant concentration (10 or 15%) and culture age used in this study, 7-day old culture appeared to be more resistant to freezing without cryoprotectant when compared with 3- and 14-day old culture. The highest cell survival values obtained with propane-1,2-diol (10%) and DMSO (10%) protection were 96.2   1.2% and 94.0   3.1% respectively. Initial subsequent cell growth after cryopreservation was slower than their non-frozen controls. The survival of transfected BF-2 cells (BF-2/luc1) after cryopreservation were very similar to those obtained with wild type cells being: 94.0   3.1% and 95.2   1.5% respectively with 10% DMSO protection. These results suggested that genetically modified fish cell lines may be equally amenable to cryopreservation as the wild type.
Zhang
Tiantian
T
Luton Institute of Research in the Applied Natural Sciences, University of Luton, 2 Adelaide Street, Luton, Bedfordshire, LU1 5DU, United Kingdom.
Rawson
David M
DM
D000818:Animals; D002460:Cell Line; D002470:Cell Survival; D015925:Cryopreservation; D003451:Cryoprotective Agents / Q000633:toxicity; D005347:Fibroblasts / Q000166:cytology; D017930:Genes, Reporter; D008156:Luciferases / Q000235:genetics*; D008163:Luminescent Measurements; D010473:Perciformes; D018675:Toxicity Tests; D014162:Transfection
D003451:Cryoprotective Agents; D008156:Luciferases