Keywords: down sides cryopreservation, advantages cryopreservation
In the world of knowledge this two methods Vitrification and Poor cooling are use to keep carefully the biological materials such as cells, bone marrow, DNA etc at the reduced temperature, in comparison with their normal temps. These two strategies will come under the Cryobiology.
Cryobiology:
It is the analysis of life below the low temperature.
Background:
In the generations 2500 BC this is employed by the people of Egypt for the medical purpose. They used to stop the blood loss and injuries through the injuries. Inside the latter centuries this was helped bring into popular by Robert Boyle. For the first time it was the Christopher Polge who used the bull sperm in cryopreservation. The 1970's brought great development in cryobiology by Zeo Layland who helped bring Slow Cooling technique which laid a way to the beginning of first individuals embryo frozen, which latter used around the globe for the family pets, cells and human biology. In the year 1986 Dr. Christopher Chen in Australia used the sluggish iced oocytes for the pregnancy on earth for the first time.
Advantages of cryobiology:
- Helps in the preservation of natural materials.
- By this the natural materials can be conserved for long time.
- Sperm, gametes, embryos, tissues, bone marrow, organ can be conserved.
- Helps to study the adapting characteristics of plants and animals under the reduced temperature.
Cryopreservation:
This is the procedure, which come under the Cryobiology. This is the process where the cell is maintained under the very low temperature which causes the cell to stop its biological chemical reactions and lastly the cell contributes to fatality. But sometimes the cell which is maintained under the procedure of cryopreservation gets damage, when it is taken to the reduced temperature. A number of the natural materials are kept under suprisingly low heat range which is the liquid phase of the liquid nitrogen. Since it is the foremost process for the preservation some sophisticated biological materials which lead to avoid their biological chemical reactions. In order to be free from the chance the most two techniques used will be the Slow Cooling and Vitrification.
Background:
James Lovelock is the key person who made the Gaia theory fame. Applying this theory he said that the damage occurring to the red bloodstream cells is due to the osmotic stress during the process of the freezing. In the early many years of 1950s he said that whenever the cell encounters the increase of sodium focus make it to dehydrate for the loss of drinking water to the exterior ice which may cause the harm of the cell. In the year of 1950s these are swift development of the freezing techniques which made being able to help in delivering the pregnancies. Before this the insemination of iced sperm was helped bring into live. Last mentioned in the 1957 the scientist of the United Kingdom started out the cryopreserving the fowl sperm.
In the year of 2000s the infant was born by the cryopreservation egg, Laina Beasley created in July 2005. Not merely in the humans, this is helped bring into the family pets which made to the consequence of A Ocelots kitten delivered in Cincinnati Zoo in 2001. As freezing destruction in the cells are of two aspects. The primary is that cell gets harm due to the snow crystal, and the second reason is the damage of cell when more snow is formed because of the focus of the solute. Last mentioned in america they made a solution for this aspects of the harm in cell by the typical rate of cooling 1C/min but this rate of chilling depends on how big is the cell and this particular content in the cell. In this they are a form of anti-freeze known as the cryprotectant which is utilized to equalize the physical best parameter osmotic. Cryoprotectants have ability to protect the cell to handle the freezing personal injury which was discovered accidentally.
Cryoprotectants:
When the biological materials are stored under the preservation these are have to be protect for the long time. At same time the secured material can function for years when they are rewarmed to the sub zero level. During the process of the preservation some chemicals are used to protect them in low heat range and just as they are simply rewarmed, and really should have the ability to function for a long period. But in some instances of preservation chemicals aren't used such as in fungi, fungus. The cryoprotectants are being used in this conditions, now a day's some chemicals like dimethyl sulfoxide, glycerol. But in some of the specimens the dimethyl sulfoxide influences the preservation due to the toxicity character. (Smith, 1983) This toxicity can be reduces for some level by use of glucose
Advantages of Cryoprotectants:
- Helps the materials from speedy cooling
- Prevents from formation of glaciers in the intracellular region.
- When the cell goes through high concentration of solute it helps to avoid from dehydration (Mazur, 1984).
- Helps the cell to function even after the rewarming.
Slow air conditioning:
This is the early technique found in the cryopreservation which is used to avoid to the cell from the damage in the freezing
Background:
It is the control rate strategy which was developed in the 1970s which includes been empowered the first human being embryo birth. From then this system is used around the globe for the biological materials. And some machines which are being used in the cryopreservation bring the cell to the freezing point such as the liquid phase of the liquid nitrogen. This system machines are used to freeze the oocyte, blood products, sperms, skin, embryo, general tissue and stem cells preservation in research labs, nursing homes all around the globe. But in the slow air conditioning the cell gets dehydrate
Vitrification:
This is the new technique found in the cryopreservation which is used to avoid to the cell from the damage in the freezing. It is the preservation at extremely low temp with no freezing. In this technique can be done without the engagement of the cryoprotectants.
Background:
Right from the introduction of the slow cooling the glycerol can be used to cryobiology as the cryoprotectant for the bull sperm and blood cells. But nonetheless it is know that glycerol is not beneficial to prevent the complete organ from the damage. For more suitable cryoprotectants in those situations lots of the biotech companies functioned to build up. In the 21st century the kidney of rabbit is maintained at -135oC, which made as the vitrification cocktail, because second option the kidney which is conserved at the -135oC was again planted back to your body of rabbit, the kidney was found to be performing without any inability. At present preservation of the mind is under the improvement, they are looking to prevent the brain from damaging such as harm to the cells and loss of the storage in the mind that was encoded.
The Institute of Cryonics will work to preserve the complete body without harm in the skin cells, tissue and everything the organs that ought to again function properly when they are transplanted, this is in the improvement. Within this the freezing includes in ice crystal development, which lead to the damage of the very sensitive structures like the arteries. For a successful vitrification it requires combinations of both factors, you are the high focus of solutes in the bathing medium with the capacity of glass creation, and the other is the extreme rapid chilling of the samples. In the entire year 1985 for the first time the cryopreservation of mouse embryos by Rall and Fahy. Steps that to be used for the successful vitrification are
- concentration and structure of the vitrification solution
- The process used to equilibrate cells in this solution
- The cooling/warming conditions
- The process used to dilute skin cells from the vitrification solution
Freezing accidental injuries:
In a full time income cell the water water is most important to maintain its structure and function, when this cell is held in the freezing preservation, because of the low heat range then to its success then the cell encounters the freezing accidental injuries which may lead the living cell to devastation. When the cell is under the preservation the injury that impact is shown in the number the inverted 'U' in this the position of the cell which it can function normally is shown as the success point, when this cell is placed on to the freezing beyond its limit, that is clearly a cell has its capacity for a certain limit of low heat range or temperature, when this cell exceeds the limit of low heat the solution throughout the cell helps it be injury, in such instances the intracellular snow creation will be took place, at this time the cell causes the damage and devastation occurs. In some instances like the high chilling rate the cell undergoes both extracellular and intracellular.
Freezing traumas at high air conditioning rate:
When we take the majority of the cells they have got the thermodynamic point at -0. 5oC. However when we need to maintain the cell the cell must get freeze, to get this done the cell will be undertaken below -5oC. At this position the cell undergoes the super cooling down of which the medium about the cell and the cell remain unfrozen, due to the defensive solute that is bounded around and within the cell.
The cell which is taken up to the low heat range between your -5oC and -15oC the snow varieties in the exterior medium. At which the cell content stay super cooling in an unfrozen state. The glaciers which is created in the external medium will have an impact on the extracellular solute. The perfect solution is awareness in the extracellular solution increase when the heat gets lowers and the glaciers will be harvested, this increase of glaciers is the ice phase. Due to this the substance imbalance is happened between the natural material and the unfrozen exterior solution.
The external part of the cell gets iced when this moves off, this occurs when the bigger chemical potential then your water of the partially frozen solution beyond your cell. And this following physical event in the cell depends upon the rate of cooling in the cell. When the cooling down is sufficiently gradual, the loss of water quickly by exosmosis. When this occurs the result of the cell will dehydrate and will not freeze intracellular. TZ p3
When the cooling is too rapid the rate of which the chemical potential of normal water extracellular solution decreases is a lot faster than to the rate which water can be diffuse out of the cell and they will be the end bring about the intracellular ice formation. In the shown number the cell under the preservation will possess the outflow of the intracellular normal water which may lead to reduce of the cell and the extracellular glaciers will be shaped which leads to the shrunken cell with little or no ice produced internally. It is the indirect assumption that the forming of the snow inside the cell is unpreventable. At present many of the studies have been recommended that intracellular glaciers formation during the process of the freezing causes the loss of life or destruction of the cell. In the process of the intracellular snow formation these are three possible ways which it can be occurred.
Chilling injury:
Chilling damage is thought as the low heat range stress on the lack of freezing. Actually the term chilling injuries is used in the botany, in the early 18th centuries to describe the plants that are subjected to the reduced temperature that is chilling temp above the 0oC were often ruined irreversibly. The temps shock was initially found in 1934 showing the irreversibly damage to mammalian sperms that happened when these cell undergo rapid chilling below your body temperature at which few degree fall down rapidly ina moment of their time. At these both sperm cells and the vegetable cells the chilling injury are in the same way related mechanism. In the process of chilling incidents they are really two type's immediate chilling damage and the indirect chilling damage.
Direct chilling injury:
This is also known as the Cold great shock. This is mainly used to spell it out both phenomena, which is expressed quickly upon decrease in temperature and Reliant on cooling rate. Wintry shock injury is nearly in addition to the rate of warming. Accident is increased as the time incubation at the reduced temps is lengthened.
Indirect chilling accidents:
Indirect chilling traumas are usually evident carrying out a relatively long publicity period at the time of the reduced heat, and its allow to the independent of the rate of cooling down.
Metabolic and enzymatic techniques can find in the fast growing embryos. Especially in Drosophila and zebrafish the personal injury get more rapid at the low temperatures. That is because of the co-ordination is increased lost with decreasing heat range. The reduction in temperature will impact the enzyme rate reaction to a different degree.
SIMILARITIES BETWEEN Operated SLOW Cooling down AND VITRIFICATION: (Baudot et al. , 2002)
In the procedure of preservation both techniques hold the similarity of freezing during the procedure for preservation.
In slow cooling down the cooling is performed intracellular and extracellular and just as in vitrification, but little change at place where snow crystal creation is happened in slow cooling down and not in vitrification
Somehow both techniques are similar with slight changes through the procedure for the preservation of natural materials.
DIFFERENCES BETWEEN Operated SLOW Cooling down AND VITRIFICATION IN CRYOPRESERVATION OF BIOLOGICAL MATERIALS: (Kuleshova, 2002)
Vitrification techinque
Slow air conditioning technique
This is simple technique
This is complicated technique
This safer technique
This is high-risk technique
This more costly technique
This cheaper compare to vitrification
Ice crystal don't form along the way of freezing
In this snow crystals formation is seen
This is most successful technique
Not much success then vitrification
Cell death won't occur
Have the probabilities to the cell death
ADVANTAGES OF VITRIFICATION AND CONTROLLED SLOW COOLING IN CRYOPRESERVATION OF BIOLOGICAL MATERIALS:
In the cryopreservation the both techniques vitrification and handled cooling techniques are being used to protect the natural materials for a long time. Vitrification technique gets the uniqueness for the preservation of the oocytes, because the oocytes brought under this technique have more competent to the fertilization. This oocytes lead to the normal pregnancy. In procedure for the vitrification the glaciers crystal formation is not occurred both in the intracellular and the extracellular. In vitrification the whole cell including the medium solidify (freeze). Along the way of vitrification the cell doesn't get any destruction and don't lead the cell to death (Kasa, 2004). The preservation of materials at a manipulated slow chilling, we can store the materials at -196oC, best example is safe-keeping of hematopoietic cells (Hill et al. , 1972). The main advantages of air conditioning and warming rates are that it contains very less amount of cryoprotectants, with this it can reduce poisonous result and also osmotic personal injury (Orief et al. , 2005).
DISADVANTAGES OF Handled SLOW COOLING AND VITRIFICATION IN CRYOPRESERVATION OF BIOLOGICAL MATERILS:
When we come to the vitrification we don't face any unfavarable conditions during the procedure for preservation, because of cryoprotectants which toxic in dynamics plus more cost(Chi, 2001). Ice crystals are happened in the intracellular and the excess mobile region of cell in the process of preservation in slow-moving cooling technique. This is actually the major downside in controlled slow-moving cooling down. (Kasa, 2004).
Main Outcome Solution:
As per the reported quantity of pregnancies done after copy of embryos which were cryopreserved by vitrification. Both sluggish cooling down and vitrification strategies have successful cryopreservation of human embryos and oocytes. Both steps have healthy births, but sluggish air conditioning of oocytes offers suprisingly low success rates. Vitrification is a appealing novel approach in reproductive technology
CONCLUSION:
As per the guide and my knowledge managed slow cooling and as well as vitrification are of help approaches for the preservation of natural materials, when compared vitrification strategy is more useful way of the preservation as slow-moving cooling approach. Vitrification is a straightforward procedure that will require less time, safer and more cost effective than slow cooling.
