Advantages and Disadvantages of Cryopreservation

Advantages and Disadvantages of Cryo conservationIn the human of scholarship this two procedures Vitrification and S pathetic cooling ar mathematical proceed to keep the biologic materials such as stallphones, b unity marrow , DNA etc at the imprint temperature , when compargond to their form temperatures. These two procedures forget come chthonian the Cryobiology.CryobiologyIt is the study of bread and butter below the low temperature.BackgroundIn the centuries 2500 BC this was utilize by the people of Egypt for the medical purpose. They utilize to stop the bleeding and injuries during the injuries. In the latter(prenominal) centuries this was brought into popular by Robert Boyle. For the introductory epoch it was the Christopher Polge who utilise the prick spermatozoon in cryo deliverance. The seventies brought great development in cryobiology by Zeo Layland who brought dimmed Cooling proficiency which laid a path to the birth of first human embryo frozen, whic h latter used all(prenominal) everyplace the world for the animals, mobile phones and human biology. In the year 1986 Dr. Christopher Chen in Australia used the unbend frozen oocytes for the pregnancy in the world for the first time.Advantages of cryobiologyHelps in the saving of biologic materials.By this the biological materials butt be carry throughd for yen time.Sperm, gametes, embryos, tissues, bone marrow, organ can be preserved.Helps to study the adapting nature of plants and animals infra the low temperature.CryopreservationThis is the regale, which come under the Cryobiology. This is the work at in which the cell is unplowed under the very low temperature which causes the cell to stop its biological chemic reactions and finally the cell fall outs to death. But al nightimes the cell which is kept under the process of cryopreservation may get aggrieve, when it is pull backn to the low temperature. Some of the biological materials atomic number 18 kept under very low temperature which is the fluidness flesh of the liquid nitrogen. Because it is the best process for the preservation some(a) complex biological compounds which lead to stop their biological chemic reactions. In order to be free from the risk the most two proficiencys used be the sluggish Cooling and Vitrification.BackgroundJames Lovelock is the important person who made the germanium theory fame. Using this theory he said that the damage that occurs to the red air cells is due to the osmotic stress during the process of the halt. In the premature years of 1950s he said that when the cell faces the maturation of season submerging make it to dehyd aim for the loss of water to the outdoor(a) trash which may cause the damage of the cell. In the year of 1950s they are quick development of the halt techniques which made helping in manner of speaking the pregnancies. Before this the insemination of frozen sperm was brought into live. Latter in the 1957 the scienti st of the United body politic started the cryopreserving the fowl sperm.In the year of 2000s the baby was innate(p) by the cryopreservation egg, Laina Beasley born in July 2005. Not only in the human beings, this is brought into the animals which made to the moment of A Ocelots kitten born in Cincinnati Zoo in 2001. As frost damage in the cells are of two aspects. The primary one is that cell gets damage due to the folderol crystal, and the second is the damage of cell when more than ice is formed due to the concent aim of the solute. Latter in the USA they made a answer for this aspects of the damage in cell by the typical pasture of cooling 1C/min but this browse of cooling depends on the size of the cell and the water content in the cell. In this they are a form of anti-freeze know as the cry entertainant which is used to equalize the physical optimal parameter osmotic. Cryo cheerants deport ability to protect the cell to face the freezing detriment which was discovered accidentally.CryoprotectantsWhen the biological materials are kept under the preservation they are shoot to be protect for the long time. At same time the protected material should be able to function for a long time when they are rewarmed to the shade zero level. During the process of the preservation some chemicals are used to preserve them in low temperature and in the same way they are rewarmed, and should meet the ability to function for a long time. But in some cases of preservation chemicals are non used such as in fungi, yeast. The cryoprotectants are used in this cases, now a days some chemicals like dimethyl sulfoxide, glycerol. But in some of the specimens the dimethyl sulfoxide affects the preservation due to the toxicity nature. (Smith, 1983) This toxicity can be reduces to some level by use of glucoseAdvantages of CryoprotectantsHelps the material from rapid coolingPrevents from establishment of ice in the intracellular region.When the cell undergoes juicy conce ntration of solute it helps to stay from dehydration (Mazur, 1984).Helps the cell to function even after the rewarming.Slow coolingThis is the former(a) technique used in the cryopreservation which is used to prevent to the cell from the damage in the freezingBackgroundIt is the control rate technique which was developed in the 1970s which has been enabled the first human embryo birth. From then this technique is used all over the world for the biological materials. And some machines which are used in the cryopreservation bring the cell to the freezing point such as the liquid phase of the liquid nitrogen. This technique machines are used to freeze the oocyte, blood products, sperms, skin, embryo, general tissues and stem cells preservation in research labs, hospitals all over the world. But in the easy cooling the cell gets dehydrateVitrificationThis is the pertly technique used in the cryopreservation which is used to prevent to the cell from the damage in the freezing. It is t he preservation at extremely low temperature without each freezing. In this process can be done without the involvement of the cryoprotectants.Background decently from the development of the lento cooling the glycerol is used to cryobiology as the cryoprotectant for the bull sperm and blood cells. But however it is know that glycerol is non helpful to prevent the undivided organ from the damage. For more suitable cryoprotectants in those cases many of the biotech companies worked to develop. In the 21st century the kidney of rabbit is preserved at -135oC, which made as the vitrification cocktail, because latter the kidney which is preserved at the -135oC was over again set back into the body of rabbit, the kidney was found to be functioning without any failure. At present preservation of the brain is under the progress, they are looking to prevent the brain from damaging such as damage to the tissues and loss of the keeping in the brain which was encoded.The Institute of Cryo nics are working to preserve the whole body without damage in the cells, tissue and all the organs which should again function properly when they are transplanted, this is in the progress. In this the freezing involves in ice crystal formation, which lead to the damage of the sensitive structures such as the blood vessels. For a successful vitrification it needs combinations of the two factors, one is the high concentration of solutes in the bathing medium capable of glass formation, and the other is the extreme rapid cooling of the samples. In the year 1985 for the first time the cryopreservation of mouse embryos by Rall and Fahy. Steps that to be followed for the successful vitrification areconcentration and composition of the vitrification solutionThe procedure used to equilibrate cells in this solutionThe cooling/warming conditionsThe procedure used to dilute cells from the vitrification solutionfrost injuriesIn a living cell the liquid water is most important to master(prenom inal)tain its structure and function, when this cell is kept in the freezing preservation, due to the low temperature then to its survival of the fittest then the cell faces the freezing injuries which may lead the living cell to destruction. When the cell is under the preservation the imperfection that effect is shown in the figure the inverted U in this the position of the cell which it can function normally is shown as the survival point , when this cell is put on to the freezing beyond its limit, that is a cell has its own capability for a certain limit of low temperature or high temperature, when this cell exceeds the limit of low temperature the solution near the cell makes it injury, in such cases the intracellular ice formation pull up stakes be occurred, at this stage the cell leads to the injury and destruction occurs. In some cases like the high cooling rate the cell undergoes twain the extracellular and intracellular.Freezing injuries at high cooling rateWhen we tak e most of the cells they have the thermodynamic point at -0.5oC. But when we need to preserve the cell the cell must get freeze, to do this the cell allow for be underinterpreted below -5oC. At this position the cell undergoes the highly cooling at which the medium around the cell and the cell lodge unfrozen, due to the protective solute that is bounded around and within the cell.The cell which is taken to the low temperature between the -5oC and -15oC the ice forms in the external medium. At which the cell content remain super cooling in an unfrozen state. The ice which is formed in the external medium will affect the extracellular solute. The solution concentration in the extracellular solution will increase when the temperature gets decreases and the ice will be grown, this increase of ice is the ice phase. overdue to this the chemical imbalance is occurred between the biological material and the unfrozen external solution.The external part of the cell gets frozen when the wa ter flows off, this occurs when the higher chemical potential then the water of the partly frozen solution orthogonal the cell. And this subsequent physical event in the cell depends on the rate of cooling in the cell. If the cooling is sufficiently dull, the loss of water speedily by exosmosis. When this occurs the result of the cell will dehydrate and will not freeze intracellular. TZ p3When the cooling is too rapid the rate at which the chemical potential of water extracellular solution decreases is much faster than to the rate which water can be diffuse out of the cell and they will be the end result in the intracellular ice formation. In the shown figure the cell under the preservation will have the natural spring of the intracellular water which may lead to shrink of the cell and the extracellular ice will be formed which leads to the shrunken cell with short or no ice formed internally. It is the indirect assumption that the formation of the ice inside the cell is unpreve ntable. At present many of the studies have been suggested that intracellular ice formation during the process of the freezing causes the death or damage of the cell. In the process of the intracellular ice formation they are three possible ways which it can be occurred. cast down injuriesChilling injury is defined as the low temperature stress on the absence seizure of freezing. Actually the word dispirit injuries is used in the botany, in the early 18th centuries to describe the plants which are subjected to the low temperature that is cast down temperature above the 0C were often damaged irreversibly. The temperature shock was first used in 1934 to show the irreversibly damage to mammalian sperms that occurred when these cell undergo rapid cooling below the body temperature at which few degree accrue down rapidly in a minute of time. At these two sperm cells and the plant cells the chilling injury are quasi(prenominal)ly link up mechanism. In the process of chilling injurie s they are two types direct chilling injury and the indirect chilling injury.Direct chilling injuriesThis is also known as the insensate shock. This is mostly used to describe both(prenominal) phenomena, which is uttered quickly upon reduction in temperature and Dependent on cooling rate. Cold shock injury is almost independent of the rate of warming. Injury is increase as the period incubation at the reduced temperature is extended.Indirect chilling injuriesIndirect chilling injuries are usually evident following a relatively long exposure period at the time of the reduced temperatures, and its enable to the independent of the rate of cooling.Metabolic and enzymatic processes can invent in the fast developing embryos. Especially in Drosophila and zebrafish the injury get more rapid at the low temperatures. This is due to the co-ordination is alter magnitude lost with decreasing temperature. The reduction in temperature will affect the enzyme rate reaction to a different extent .SIMILARITIES BETWEEN CONTROLLED SLOW temperature reduction AND VITRIFICATION (Baudot et al., 2002)In the process of preservation both the techniques have the similarity of freezing during the process of preservation.In slow cooling the cooling is done intracellular and extracellular and in the same way in vitrification, but little change at place where ice crystal formation is occurred in slow cooling and not in vitrificationSomehow both techniques are similar with slight changes during the process of the preservation of biological materials.DIFFERENCES BETWEEN CONTROLLED SLOW chilling AND VITRIFICATION IN CRYOPRESERVATION OF BIOLOGICAL MATERIALS (Kuleshova, 2002)Vitrification techinqueSlow cooling techniqueThis is simple techniqueThis is complex techniqueThis safer techniqueThis is risky techniqueThis more dear(p) techniqueThis cheaper compare to vitrificationIce crystal dont form in the process of freezingIn this ice crystals formation is seenThis is most successful techniqueNo t much success then vitrificationCell death will not occurHave the chances to the cell deathADVANTAGES OF VITRIFICATION AND CONTROLLED SLOW COOLING IN CRYOPRESERVATION OF BIOLOGICAL MATERIALSIn the cryopreservation the both techniques vitrification and controlled cooling techniques are used to preserve the biological materials for a long time. Vitrification technique has the uniqueness for the preservation of the oocytes, because the oocytes brought under this technique have more capable to the fertilization. This oocytes lead to the normal pregnancy. In process of the vitrification the ice crystal formation is not occurred both in the intracellular and the extracellular. In vitrification the whole cell including the medium change integrity (freeze). In the process of vitrification the cell doesnt get any damage and dont lead the cell to death (Kasa, 2004). The preservation of materials at a controlled slow cooling, we can store the materials at -196oC, best example is storage of h ematopoietic cells (Hill et al., 1972). The main advantages of cooling and warming rates are that it contains very less issue forth of cryoprotectants, with this it can reduce toxic effect and also osmotic injury (Orief et al., 2005).DISADVANTAGES OF CONTROLLED SLOW COOLING AND VITRIFICATION IN CRYOPRESERVATION OF BIOLOGICAL MATERIALSWhen we come to the vitrification we dont face any unfavarable conditions during the process of preservation, because of cryoprotectants which toxic in nature and more cost(Chi, 2001). Ice crystals are occurred in the intracellular and the extra cellular region of cell in the process of preservation in slow cooling technique. This is the major disfavor in controlled slow cooling. (Kasa, 2004).Main Outcome MeasureAs per the describe number of pregnancies done after transfer of embryos which were cryopreserved by vitrification. Both slow cooling and vitrification procedures have successful cryopreservation of human embryos and oocytes. Both procedures have ruddy births, but slow cooling of oocytes gives very low success rates. Vitrification is a promising novel technique in reproductive technology inferenceAs per the reference and my knowledge controlled slow cooling and as come up as vitrification are useful techniques for the preservation of biological materials, when compared vitrification technique is more useful technique for the preservation as slow cooling technique. Vitrification is a simple procedure that requires less time, safer and more cost effective than slow cooling.