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Evaluation of different methods of cryopreservation of Ehrlich tumor cells
Kamilla M Santos1, Fernanda P S Maciel1, Cristina M Souza2
1Federal University of Sao Joao del Rei-CCO, Laboratory of Experimental Pathology, Divinopolis, Minas Gerais, Brazil.
Background:
The Ehrlich Ascitic Carcinoma (EAC) is an experimental transplantable neoplasm that develops in several species of mice. The maintenance of the tumor occurs in vivo. Thus, freezing the cells would reduce the number of passages between animals, ensuring genetic stability and storage for longs period of experimentation.
Objective:
Search by EAC cryoprotectants.
Materials And Methods:
The combinations of nutrient medium (Tris, hen egg yolk, and DEMEN) and cryoprotective agent (Glicerol, Trehalose and DMSO) on freezing EAC cells and the transplantability after defrosting were evaluated. The cooling was conducted at 2 C/min. until -180 degree C and the thawing by immersion in water at 37 degree C. The transplantability was evaluated from cell inoculation in mice for 14 days.
Results:
The best results were the associations IA (Cryoprotective agent Glycerol 6 % and medium containing 3.0 % Tris w / v, 1.8 % Citric acid w / v, 1.3 % D-fructose w / v and 20 % hen egg yolk v / v) and IIB (Cryoprotective agent Trehalose 100mM and medium containing 50 % coconut water v / v, 25 % sodium citrate 5 % v / v and 20 % hen egg yolk v / v) with 85.2 % and 55.1 % viable cells, respectively.
Conclusion:
These transplantable cells were efficient for tumor development, therefore demonstrating that this method of cryopreservation is simple and affordable.
