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CryoLetters 22, 299-310 (2001)
CryoLetters, c/o Royal Veterinary College, London NW1 OTU, UK

CRYOPRESERVATION OF PRIMARY CELL CULTURES OF
MARINE INVERTEBRATES

Nelly Odintsova*, Konstantin Kiselev**, Nina Sanina** and Edward Kostetsky**

*Institute of Marine Biology, Far East Branch of Russian Academy of Sciences, Vladivostok, Russia 690041.
**Far Eastern State University, Vladivostok, Russia 690090. E-mail: kkv5@mail.ru.


Abstract
Primary cell cultures obtained from somatic and larval tissues of bivalve molluscs and from embryos of sea urchins were frozen to -196oC by two-step freezing using 10% dimethyl sulfoxide (DMSO) or/and trehalose (3-30mg/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.

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