CRYOPRESERVATION OF PECTEN-MAXIMUS HEART-CELLS

A dissociation protocol for Pecten maximus heart has been established that makes it possible to obtain functional primary cultures routinely (9, 10); freezing assays of isolated cells were performed with the ai m of making it possible to cultivate these cells in vitro after thawin g to provide a constant standardized source of cells for applied resea rch. Various parameters such as the nature and the concentration of th e cryoprotectant, the cooling rate, and the incubation time of cells w ith the cryoprotective agent were evaluated. Best results were obtaine d by freezing cells in 12% dimethyl sulfoxide (Me2SO) in Leibovitz L15 medium at a cooling rate of approximately 2-3 degrees C/min. Thawed c ells in culture attached to the substrate and survived for at least 3 weeks. They exhibited similar morphology and synthesised proteins, DNA , and lipids bl vitro at levels close to those observed in fresh cells . To our knowledge, it is the first time that cultures have been obtai ned from cryopreserved marine invertebrate cells, (C) 1998 Academic Pr ess.