Importance of Cryopreservation, Methods of storage, Advantages of Cryopreservation, Stages of cryopreservation.

1. PRESENTED TO:
Dr.Koyeli Girigoswami,
Associate professor FAHS,CARE.
M.Sc. Medical Genetics & Molecular
Diagnostics
PRESENTATION BY:
R.Azmathullah
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2. Cryopreservation is a process that preserves - Organelles,
Cells, Tissues, or any other biological constructs by
cooling the samples to very low temperatures.
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CRYOPRESERVATION – BACKGROUND

3. • The word cryo comes from the Greek word "kayos" - "frost".
• It means preservation in a "frozen state".
• It is the process of cooling and storing cells, tissues, or organs at
very low temperatures to maintain their viability.
• The temperature using Solid carbon dioxide −80oC or Using
liquid nitrogen −196oC.
• In this technique, tissues can be preserved for a very long time.
The science that deals with cryopreservation is known as
“cryobiology”.
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CRYOPRESERVATION – INTRODUCTION

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IMPORTANCE OF CRYOPRESERVATION
• Fertility treatments.
• Minimal space and labour required.
• Safety from genetic contamination.
• Safeguards genetic integrity of valuable stains.
• Safeguards the germplasm of endangered species.
• Biological samples can be preserved for a longer period of
time.
• Protects the samples from disease and microbial
contamination.
• Prevents genetic drift by cryopreservation of gametes,
embryos, etc.

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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.
• A sugar called trehalose, which occurs in organisms capable of surviving
extreme dehydration, is used for freeze-drying methods of
cryopreservation.
• Trehalose stabilizes cell membranes, and it is particularly useful for the
preservation of sperm, stem cells, and blood cells.

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STEPS INVOLVED IN
CRYOPRESERVATION
Selection of Material
Addition of Cryoprotectant
Freezing
Storage in Liquid Nitrogen
Thawing
Washing & Reculturing
Measurement of Viability
Regeneration

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METHODS OF STORAGE- CRYOPRESERVATION
• Cryopreservation
• Cold storage
• Low pressure
• Low oxygen storage

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METHODS OF STORAGE
Cryopreservation Cold storage Low pressure Low oxygen storage
It generally involves
storage in liquid nitrogen It involves
storage in low
and non
freezing
temperature
it involves partially
reducing the
atmospheric
pressure of
surrounding
it involves reducing the oxygen
level but maintaining the
pressure

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 Preservation of biological materials.
 Biological materials can be kept for a long
period
 Genetic integrity of the material is stored.
 It is possible to preserve gametes, embryos,
and other essential cells or tissues.
 Study of adaptations of plants and animals
to low temperatures.

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 Once the material is successfully conserved at a
particular temperature, it can be preserved
identifiably.
 No change or contamination of fungus or
bacteria takes place after the storage process is
completed and material is preserved.
 Minimal space is required for the purpose of
cryopreservation.
 Minimal labor is required for the purpose of
cryopreservation.

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