Vitrification of caudal fin explants from zebrafish adult specimens.
No data on vitrification of tissue samples are available in fishes. Three vitrification solutions were compared: V1: 20% ethylene glycol and 20% dimethyl sulphoxide; V2: 25% propylene glycol and 20% dimethyl sulphoxide, and; V3: 20% propylene glycol and 13% methanol, all three prepared in Hanks' buffered salt solution plus 20 percent FBS, following the same one step vitrification procedure developed in mammals. Caudal fin tissue pieces were vitrified into 0.25 ml plastic straws in 30s and stored in liquid nitrogen for 3 days minimum, warmed (10s in nitrogen vapour and 5s in a 25 degree C water bath) and cultured (L-15 plus 20% FBS at 28.5 degree C). At the third day of culture, both attachment and outgrowing rates were recorded. V3 led to the worst results (8% of attachment rate). V1 and V2 allow higher attachment rates (V1: 63% vs V2: 50%. P < 0.05) but not significantly different outgrowing rates (83% to 94%). Vitrification of caudal fin pieces is advantageous in fish biodiversity conservation, particularly in the wild, due to the simplicity of procedure and equipment.
Cardona-Costa
J
J
Laboratory of Animal Reproduction and Biotechnology (LARB-UPV), Polytechnic University of Valencia, Valencia, Spain. cardona_costa_j@hotmail.com
Roig
J
J
Perez-Camps
M
M
García-Ximénez
F
F
D000818:Animals; D015925:Cryopreservation / Q000379:methods*; D003451:Cryoprotective Agents / Q000494:pharmacology / Q000592:standards; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D004338:Drug Combinations; D019855:Ethylene Glycol / Q000494:pharmacology; D000432:Methanol / Q000494:pharmacology; D009924:Organ Culture Techniques; D019946:Propylene Glycol / Q000494:pharmacology; D015027:Zebrafish
D003451:Cryoprotective Agents; D004338:Drug Combinations; D019946:Propylene Glycol; D019855:Ethylene Glycol; D000432:Methanol; D004121:Dimethyl Sulfoxide