Cryopreservation of goldfish fins and optimization for field scale cryobanking.
When gametes and embryos are not available, cryobanking of somatic tissues is one possibility to keep a genetic record of fish valuables in a context of biodiversity conservation and animal breeding management. Cryopreservation of whole fin pieces would be more advantageous than the commonly used cryopreservation of cells after fin culture, as it would allow extensive sampling without immediate need for laboratory facilities. The objective of this work was to assess the cryopreservation ability of fin pieces from goldfish (Carassius auratus) and to test whether a laboratory procedure could be adapted to field conditions. Caudal fin explants were cryopreserved in culture medium with 125mM sucrose and 10% Me(2)SO. After 14days of culture, the frozen-thawed explants showed the same cell growth rate and grew the same somatic cell number as the fresh ones. Cells proliferated inside and around the explants as shown by BrdU labeling. Neither the size of the fin pieces nor the freezer type, -70 degrees C upright or -20 degrees C chest, influenced the outcome of cryopreservation. Fin pieces were stored 4days at 4 degrees C in dry conditions prior to cryopreservation without alteration of the fin explant culture success. This study demonstrated that field collecting of goldfish fin pieces is possible as whole fin pieces can be stored in standard fridge or be shipped at subzero temperature before they are frozen into a plain -20 degrees C chest freezer. After incorporation in cryobanks in liquid nitrogen, thawed fin pieces reliably produce somatic cells in cell culture conditions.
Moritz
Charlotte
C
INRA, UR 1037 SCRIBE, Campus de Beaulieu, F-35000 Rennes, France.
Labbe
Catherine
C
D000818:Animals; D049109:Cell Proliferation; D002478:Cells, Cultured; D015925:Cryopreservation / Q000379:methods*; D003451:Cryoprotective Agents / Q000737:chemistry; D003470:Culture Media / Q000737:chemistry; D006054:Goldfish / Q000033:anatomy & histology*; D012034:Refrigeration; D014022:Tissue Survival
D003451:Cryoprotective Agents; D003470:Culture Media