Cryopreservation of mantle dissociated cells from Haliotis tuberculata (Gastropoda) and postthawed primary cell cultures

• Published in: Cryobiology Vol. 44; no. 1; pp. 38 - 45
• Main Authors: Poncet, J.-M, Serpentini, A, Boucaud-Camou, E, Lebel, J.-M
• Format: Journal Article
• Language: English
• Published: San Diego, CA Elsevier Inc 01.02.2002; Elsevier
• Subjects: Animal biology; Animals; Biochemistry. Physiology. Immunology; Biological and medical sciences; Cell Separation; Cell Survival; Cells, Cultured; Cryopreservation; Cryopreservation - methods; Cryoprotective Agents; Dimethyl Sulfoxide; Fundamental and applied biological sciences. Psychology; Glycerol; Haliotis tuberculata; Insulin - pharmacology; Insulin-Like Growth Factor I - pharmacology; Invertebrate Zoology; Invertebrates; Life Sciences; Mantle cells; Mollusc; Mollusca; Mollusca - cytology; Mollusca - metabolism; Physiology. Development; Protein Biosynthesis; Cryopreservation; Mollusc; Mantle cells; Laboratory study; Thawing; Gastropoda; Metabolism; Freezing; Marine environment; Enzymatic activity; Haliotis tuberculata; Animal; Primary culture; Invertebrata; Mollusca; Mantle; Viability
• ISSN: 0011-2240; 1090-2392
• DOI: 10.1016/S0011-2240(02)00001-9

Dissociated mantle cells from the gastropod mollusc Haliotis tuberculata were cultured after a freezing–thawing procedure using either 10% dimethyl sulfoxide (Me 2SO) or 10% glycerol (Gly) as a cryoprotector. The survival rate of 2-day-old cultured cryopreserved cells after thawing, based on analysis of DNA and protein contents, was nearly 80% in comparison with 2-day-old cultured fresh cells. Cells thawed after cryopreservation exhibited the maintenance of all tested physiological activities. Metabolic activity (measured by the MTT test) and the activity of alkaline phosphatase (a plasma membrane-bound enzyme) were not decreased in comparison to those in cultured fresh cells. In addition, cryopreserved cultured cells maintained a physiological stimulation ability in response to treatment with growth factors. These results taken together represent one of the most convincing demonstrations of the survival and of the recovery of intact functional activities of molluscan cells after a freeze–thawing procedure. Our results suggest that in the future primary cultures of cryopreserved mantle cells will be able to be used for fundamental research, in toxicity tests, or in the field of biotechnology.