Cryobanking of fish somatic cells: optimizations of fin explant culture and fin cell cryopreservation.
When gametes or embryos are not available, somatic cells should be considered for fish genome cryobanking of valuable or endangered fish. The objective of this work was to develop a method for fin explant culture with an assessed reliability, and to assess fin cells ability to cryopreservation. Anal fins from goldfish (Carassius auratus) were minced and gently loosened with collagenase before explants were plated at 20 degrees C in L-15 medium supplemented with fetal bovine serum and pH buffering additives. Quantification of cell-donor explants per fin rated the culture success. Cells were successfully obtained from every cultured anal fin (mean = 65% cell-donor explant per fin). All other fin types were suitable except the dorsal fin. Explant plating could be deferred 3 days from fin collecting. Fins from seven other fish species were successfully cultured with the method. After 2-3 weeks, sub-confluent fin cells from goldfish were cryopreserved. Cryopreservation with dimethyl sulfoxide and sucrose at a slow freezing rate allowed the recovery of half the goldfish fin cells. Cells displayed the same viability as fresh ones. 1,2-propanediol was unsuitable when a fast freezing rate was used. The procedure could now be considered for cryobanking with only minimal adaptation to each new species.
Mauger
P-E
PE
Group for cell cryopreservation and fish reconstruction, IFR 140, INRA-SCRIBE, Campus de Beaulieu, 35042 Rennes Cedex, France.
Le Bail
P-Y
PY
Labbé
C
C
D000818:Animals; D002470:Cell Survival; D002478:Cells, Cultured; D017364:Collagenases / Q000378:metabolism; D015925:Cryopreservation / Q000379:methods*; D003451:Cryoprotective Agents / Q000494:pharmacology; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D005399:Fishes / Q000254:growth & development* / Q000378:metabolism; D005615:Freezing; D046509:Tissue Culture Techniques
D003451:Cryoprotective Agents; D017364:Collagenases; D004121:Dimethyl Sulfoxide