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CRYOPRESERV
ATION
CRYOPRESERVATION
⚫FirstdocumentedbyBoyle(1663).
⚫Firstconceptofspermpreservationby
Lazzarospallanzini(1760).
⚫ Most significantdiscoverywastheuseof
glycerolascryoprotectant byPolgeand
smith(1949)
PRINCIPLE
1. FREEZING
2. STORING
3. THAWING
CONT
..
⚫ Theabilityofglycerol toprotectcellsfromfreezing
injurywasdiscoveredaccidentally
.
⚫ Thesubsequentdevelopmentofcryopreservation
techniqueshashad ahuge impactinmanyfields,
mostnotablyinreproductivemedicine.
⚫
⚫ Freezing injuryhasbeenshownto havetwo
components,direct damage fromtheicecrystals
andsecondarydamage causedbytheincreasein
concentrationofsolutesasprogressivelymoreiceis
formed.
⚫ Intracellularfreezingisgenerallylethalbutcanbe
avoided bysufficientlyslow cooling, and under
usualconditionssolutedamagedominates.
WHY
CRYOPRESERVATION?
⚫ Availability and preservation of raw biological
materials are the major challenge in the
widespread use of assisted reproduction
technology in humans and livestock.
⚫ For example, many artificial insemination
procedures rely on availability of cryopreserved
sperm.
⚫ Similarly, cryopreserved oocytes and embryos are
being increasingly used in assisted reproduction
CRYOPROTECTANTS
⚫ A cryoprotective agent is a substance added to the
freezing medium to prevent the biological material
from freezing injures that is damage due to ice
formation
⚫ In experimental biology, the conventional CPAs are
glycols and glycerol
Common cryoprotectants:
⚫ DMSO-Dimethyl sulfoxide
⚫ Ethylene glycol
⚫ Glycerol
⚫ Thepresence of cryoprotectantsin thefreezingsolution is
necessary topreventcell damage during freezing and
thawing of thesamples
⚫ Cryoprotectants are included in the solutions to freeze
thecells to lower thefreezing point
CONT
..
SPERM BANKING
⚫Sperm banking is the name for the
collection and storage of semen
⚫Semen is the fluid that contains sperm,
Sperm banking is also known as sperm
cryopreservation or semen storage

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