IDENTIFICATION OF MEDIA SUPPLEMENTS THAT IMPROVE THE VIABILITY OF PRIMARILY CELL-CULTURES OF CRASSOSTREA-GIGAS OYSTERS

Media supplements have been investigated for their influence on the vi ability of primary cell cultures from the heart of Crassostrea gigas o ysters. Soluble factors of vertebrate origin were tested, belonging to five families of supplements that had proven to increase the viabilit y of insect and mammal cell cultures. Using two-level complete factori al assays, factors and mutual interactions were screened within each f amily with a MTT reduction assay. Results pointed out the positive inf luence of hormones, growth factor, antioxidants and lipids bn the mito chondrial metabolism of oyter's heart cells. Consequently, a new conce ntrated complex supplement was developed. At 10% (v/v) final concentra tion in modified Leibovitz L-15 medium, it increases by 30% the cellul ar viability of one-week old cultures as compared with non-supplemente d medium, a similar improvement as the one obtained with 10% (v/v) fet al calf serum. Combined with fetal calf serum, this new supplement dou bles the cellular viability of one-week old cultures and allows networ ks of cardiomuscular cells to be maintained functional over three mont hs in vitro.