Cryopreservation of primary cell cultures of marine invertebrates.
Primary cell cultures obtained from somatic and larval tissues of bivalve molluscs and from embryos of sea urchins were frozen to -196 degrees C by two-step freezing using 10% dimethyl sulfoxide (DMSO) or/and trehalose (3-30 mg/ml) as cryoprotectants. We estimated both cell viability and the RNA synthetic activity after freeze-thaw. Total lipid extracts from the tissues of echinoderms examined as possible cryoprotective agents demonstrated a weak cryoprotective capacity. Mussel lipid extract was found to possess a considerable cryoprotective activity. Cryoprotective capacity of tested lipids correlated with their thermotropic behaviour. DMSO + trehalose combination was shown to be a favourable cryoprotectant and sea urchin blastula cells the most freezing-tolerant cells.
Odintsova
N
N
Institute of Marine Biology, Far East Branch of Russian Academy of Sciences, Vladivostok, Russia 690041. inmarbio@mail.primorye.ru
Kiselev
K
K
Sanina
N
N
Kostetsky
E
E
D000818:Animals; D002470:Cell Survival; D002478:Cells, Cultured; D015925:Cryopreservation; D003451:Cryoprotective Agents; D004121:Dimethyl Sulfoxide; D004625:Embryo, Nonmammalian; D005880:Gills / Q000166:cytology / Q000378:metabolism; D007814:Larva; D008974:Mollusca / Q000166:cytology*; D009132:Muscles / Q000166:cytology / Q000378:metabolism; D012313:RNA / Q000096:biosynthesis; D017694:Rewarming; D012617:Sea Urchins / Q000166:cytology*; D014199:Trehalose; D014529:Uridine / Q000378:metabolism
D003451:Cryoprotective Agents; D012313:RNA; D014199:Trehalose; D014529:Uridine; D004121:Dimethyl Sulfoxide