cryopreservation techniques in animal cell culture

1. CRYOPRESERVATION

2. CRYOPRESERVATION
• THE ABILITY TO FREEZE AND THAW WITH RETENTION OF VIABILITY
• PROVIDES FLEXIBILITY IN HUMAN INFERTILITY THERAPY WHEN GAMETES OR
EMBRYOS ARE HANDLED IN VITRO BECAUSE FROZEN TISSUE CAN BE STORED
INDEFINITELY IN LIQUID NITROGEN AT -196°C
• BOON TO ANIMAL HUSBANDRY – ART
• GAMETE / EMBRYO BANKING

3. PRINCIPLE OF CRYOPRESERVATION
• INTRACELLULAR ICE CRYSTAL FORMATION
• EXPOSURE TO HIGH SOLUTE CONC. / OSMOTIC SHOCK IN FREEZE / THAWING
• OSMOTIC CHALLENGE IN THE TEMPERATURE TRANSITION DEPENDS ON
• CELL'S PERMEABILITY COEFFICIENT,
• ITS SURFACE AREA, AND
• THE OSMOTIC GRADIENT BETWEEN OUTSIDE AND INSIDE
• CELLULAR DEHYDRATION & LOWERING OF FREEZING POINT

4. CYROSURVIVAL DEPENDS ON
• CELL TYPE,
• CRYOPROTECTANT SELECTION,
• CRYOPROTECTANT ADDITION,
• SEEDING,
• COOLING RATE,
• FINAL COOLING TEMPERATURE,
• HYDRATION STATE OF THE CELLS,
• WARMING RATE, AND
• CRYOPROTECTANT REMOVAL