Video link: https://youtu.be/XmBA4DwMWFw What is used for cryopreservation? What cryopreservation is used for? Why is cryopreservation needed? Principal of cryopreservation Natural cryopreservation Microscopic multicellular organisms called tardigrades can withstand freezing by displacing most of their internal water with the sugar trehalose, preventing it from crystallising and harming cell membranes. By cooling to shallow temperatures (typically 80 °C or 112 °F using solid carbon dioxide or 196 °C (321 °F) using liquid nitrogen), organelles, cells, tissues, extracellular matrix, organs, or any other biological constructs susceptible to damage caused by uncontrolled chemical kinetics are preserved. If the temperature is low enough, all enzymatic or chemical activity that could harm the natural material is effectively halted. The goal of cryopreservation techniques is to freeze objects at extremely low temperatures without inflicting additional harm from ice crystal growth Risk factors The following are the primary dangers of embryo cryopreservation: damage caused by freezing embryos. unfrozen embryos that are not viable. failure to conceive after thawing and implanting embryos.

1. CRYOPRESERVATION OF
CELLS AND TISSUES

2. • The preservation of cells and tissue by cooling
the sample to very low temperature.
• Two common cryoprotective agents
are dimethyl sulfoxide (DMSO) and glycerol.
Glycerol is used primarily for cryoprotection of red
blood cells, and
DMSO is used for protection of most other cells and
tissues.
WHAT IS CRYOPRESERVATION?

3. Trehalose stabilizes cell membranes, and it is particularly useful for the
preservation of sperm, stem cells, and blood cells.
Water-bears (Tardigrada), can survive freezing by replacing most of
their internal water with the sugar trehalose, preventing it from
crystallization that otherwise damages cell membranes
wood frogs can tolerate the freezing of their blood and other
tissues.
NATURAL CRYOPRESERVATION.

4. • Arctic and Antarctic insects, fish and amphibians create
cryoprotectants (antifreeze compounds and antifreeze proteins)
in their bodies to minimize freezing damage during cold winter
periods
Continued………….

5. MECHANISM
Selection of
material
1
Storage in liquid
nitrogen
4
Measurement of
viability
7
Washing and
reculturing
6
Addition of
cryoprotectant
2
Cooling
3
Regeneration of plant
8
Thawing
5

6. IMPORTANT APPLICATION OF
CRYOPRESERVATION
• In the freezing and storage of hematopoietic stem cells.
It has greatly enhanced the outcome for the
treatment of certain lymphomas and
solid tumour malignancies.
• Disease free plant can be preserved and propagated.
• Endangered species can be maintained,
• Rare germplasm and other genetic modulations can be
stored.

7. MAJOR ADVANTAGES
Minimum
labor
required
1
Once
material is
preserved,
it can be
preserved
indefinitely
3
Once
stored, no
chance of
contaminat
ion
4
Minimum
space
required
2

8. RISKS
• Phenomena which can cause damage to cells during
cryopreservation mainly occur during the freezing stage, and
include;
 solution effects,
 extracellular ice formation,
 dehydration, and
 intracellular ice formation.
• Many of these effects can be reduced by cryoprotectants. Once
the preserved material has become frozen, it is relatively safe
from further damage

9. • Bacteria and fungi can be kept short-term
(months to about a year, depending)
refrigerated,
however, cell division and metabolism is not
completely arrested and thus is not an optimal option
for long-term storage (years).
PRESERVATION OF MICROBIOLOGY
CULTURES