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DESIGN OF A SIMPLE SLOW COOLING DEVICE FOR CRYOPRESERVATION OF SMALL BIOLOGICAL SAMPLES
Leonardo Juan de Paz1, Maria Celeste Robert2, Daniel Adolfo Graf1
1Centro Binacional (Argentina -Italia) de Investigaciones en Criobiologia Clinica y Aplicada (CAIC); Servicio de Electronica y Optica, Facultad de Ciencias Bioquimicas y Farmaceuticas, Universidad Nacional de Rosario, Rosario, Argentina.
Background:
Slow cooling is a cryopreservation methodology where samples are cooled to its storage temperature at controlled cooling rates.
Objective:
Design, construction and evaluation of a simple and low cost device for slow cooling of small biological samples.
Materials And Methods:
The device was constructed based on Pye's freezer idea. A Dewar flask filled with liquid nitrogen was used as heat sink and a methanol bath containing the sample was cooled at constant rates using copper bars as heat conductor.
Results:
Sample temperature may be lowered at controlled cooling rate (ranging from 0.4°C/min to 6.0°C/min) down to ~-60°C, where it could be conserved at lower temperatures. An example involving the cryopreservation of Neuro-2A cell line showed a marked influence of cooling rate over post preservation cell viability with optimal values between 2.6 and 4.6°C/min.
Conclusion:
The cooling device proved to be a valuable alternative to more expensive systems allowing the assessment of different cooling rates to evaluate the optimal condition for cryopreservation of such samples.
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