Cryopreservation is the use of very low temperatures to preserve structurally intact living cells and tissues. Unprotected freezing is normally lethal and this chapter seeks to analyze some of the mechanisms involved and to show how cooling can be used to produce stable conditions that preserve life. The biological effects of cooling are ... Cryoprotectants play a vital role in the cryopreservation process, protecting biological samples from freezing damage. Here, we evaluate the effects of the combination and interaction of different extenders with permeable and non-permeable cryoprotectants, on the cryopreservation of Danio rerio sperm, analyzing the effects of cryopreservation …Aug 10, 2023 · Cryopreservation describes methods to preserve living organisms and biological materials at very low temperatures. Central to this process is the use of liquid nitrogen, enabling successful cryogenic freezing and cryopreservation, opening possibilities for cell banking and other types of ex vivo preservation over extended periods of time. Cryopreservation involves storing biological material at ultra-low temperatures, usually in liquid nitrogen. This allows long-term preservation by stopping almost all metabolic activity in cells. Materials are frozen using slow freezing, rapid freezing, or stepwise freezing methods. They are then stored long-term at temperatures near -196°C.The impact of cryopreservation on the sperm membrane is well known, and many studies agree that frozen–thawed semen is less fertile than fresh semen because of reduced sperm motility and a lower number of viable spermatozoa [ 27, 28 ]; however, it does not seem to negatively influence the live birth rate [ 29 ]. 2.2.Cryopreservation is a key method of preservation of biological material for both medical treatments and conservation of endangered species. In order to avoid …Mar 24, 2021 · Cryopreservation is the storage of biological material at low temperatures. Since ancient times, it has been known that biological material can be preserved longer at low temperatures. Indeed, archaeological findings indicate that as early as 2000 BC, icehouses were used throughout Mesopotamia to store foods [ 1 ]. Cryopreservation gained prominence in human medicine after its use in infertility treatment. Since then, gamete cryopreservation has been developed to combat infertility. Sperm was the first successfully frozen reproductive cell and remains the easiest to freeze due to its tiny cytoplasm and thus low water content. Cryopreservation is a key method of preservation of biological material for both medical treatments and conservation of endangered species. In order to avoid …The objective of this study was to investigate the recovery of bacteria from ewe milk after freezing for 4 or 8 weeks with and without the addition of glycerol as a cryopreservant. A total of 50 udder-half milk samples with a known range of bacterial species were selected, stored and analyzed in 5 treatment-groups: time zero, frozen for 4 weeks with, and …Cryopreservation is a key enabling technology in regenerative medicine that provides stable and secure extended cell storage for primary tissue isolates and constructs and prepared cell preparations. The essential detail of the process as it can be applied to cell-based therapies is set out in this review, covering tissue and cell isolation, …Core Tip: The manuscript is an overview of current cryopreservation protocols used for cold storage of hematopoietic stem cells, mesenchymal stem cells and induced pluripotent stem cells.Although dimethyl sulfoxide (DMSO) is commonly used in cryopreservation of cell lines, primary cells and stem cells, the use of DMSO has been …Jul 18, 2022 · Cryopreservation, the process of storing materials at sub-zero temperatures, is an essential process for fundamental research, as well as the clinical, biomedical and food sciences 1. The ability... Impact of Cryopreservation on Viability and Recovery Rates. In the second study, PBMC viability, as assessed by trypan blue staining, was 99.2 ± 0.8 in fresh samples, while the average values of frozen PBMC replicas were 94%, 92%, 93%, 97%, and 93%. No replicas of any animal dropped below 88%.Cryopreservation is the most prevalent method of long-term cell preservation. Effective cell cryopreservation depends on freezing, adequate storage, and correct thawing …Jun 9, 2022 · Cryopreservation is performed either through programmable slow freezing, vitrification or low-CPA vitrification (ultra-rapid cooling) with vitrification being is the preferred technique in cryonics because it prevents/reduces the formation of damaging ice crystals within the cryopreserved subject . Dec 13, 2021 · Plant cryopreservation, specifically, is a process of cooling and storing vegetal structure as plant cells, tissues, or organs in liquid nitrogen (LN; −196 °C) or LN vapor (−160 °C). This methodology ensures the maintenance of samples’ viability after thawing, and indefinite storage is possible. The cryopreservation technique is based ... Abstract. Cryopreservation plays a significant role in tissue and organ banking. With the rapid development of tissue engineering, cryopreservation of tissue-engineered products is of great importance to meet the ‘ready-to-use’ requirement in clinical application and regenerative medicine. Cryopreservation can be achieved by conventional ...The cryopreservation of hematopoietic cells using dimethyl sulfoxide (DMSO) and serum is a common procedure used in transplantation. However, DMSO has clinical and biological side effects due to ...Cryopreservation is a key enabling technology in regenerative medicine that provides stable and secure extended cell storage for primary tissue isolates and …Cryopreserve definition: to maintain the viability of (cells, tissue, organs, etc.) by storing them at very low temperatures.. See examples of CRYOPRESERVE used in a sentence.Cryopreservation is the application of low temperatures to preserve the structural and functional integrity of cells and tissues. Conventional cooling protocols allow ice to form and solute concentrations to rise during the cryopreservation process. The damage caused by the rise in solute concentration can be mitigated by the use of compounds ...Jun 23, 2020 · Principle of Cryopreservation: Cryopreservation is a process of preserving or storing cells, tissues, organs or any other biological materials from any potential damage by maintaining the materials at very low temperature (typically -80 °C using solid CO2 or −196 °C using liquid Nitrogen. In cryopreservation, very low temperatures is used ... Feb 1, 2021 · 1 Introduction. Cryopreservation is a basic and important technique to achieve long-term storage of organs, tissues, cells, and other biological materials by using a very low temperature (at −80 or −196 °C). Jun 23, 2020 · Principle of Cryopreservation: Cryopreservation is a process of preserving or storing cells, tissues, organs or any other biological materials from any potential damage by maintaining the materials at very low temperature (typically -80 °C using solid CO2 or −196 °C using liquid Nitrogen. In cryopreservation, very low temperatures is used ... Apr 18, 2023 · In addition, the future of sperm cryopreservation might benefit from personalized and individually designed cryopreservation approaches. However, established cryopreservation programs with highly developed protocols and appropriate devices can better support FP procedures and may reshape the cryo-banking landscape of gametes and embryos. Cryopreservation is a critical procedure in autologous hematopoietic stem cell transplantation. Dimethyl sulfoxide (DMSO) is the cryoprotectant of choice. Optimization of the cryopreservation protocol in the past revealed a dramatic loss of cell viability associated with a reduction of the DMSO concentration below 2 vol % in the freezing … In addition to general considerations, cell-specific recommendations for hepatocytes, pancreatic islets, sperm, oocytes, and stem cells should be observed to maximize yields. For example, rapid cooling is associated with better cryopreservation outcomes for oocytes, pancreatic islets, and embryonic stem cells while slow cooling is recommended ... Cryopreservation is routinely used for the long-term storage of cells in various areas of academic, industrial, and clinical research. To keep frozen cells alive, it is necessary to vitrify (or nanocrystallize) water on the inside and outside of the cells. Vitrification is conventionally achieved by adding at least one cryoprotective agent (CPA ...Cryopreservation or cryoconservation is a process where biological material - cells, tissues, or organs - are frozen to preserve the material for an extended period of time. At low temperatures (typically −80 °C (−112 °F) or −196 °C (−321 °F) using liquid nitrogen) any cell metabolism which might cause damage to the … See moreCryopreservation is the most prevalent method of long-term cell preservation. Effective cell cryopreservation depends on freezing, adequate storage, … It is now accepted that with current methods of cryopreservation, uncontrolled ice formation in structured tissues and organs at subzero temperatures is the single most critical factor that severely restricts the extent to which tissues can survive procedures involving freezing and thawing. In recent years, this major problem has been ... Cryopreservation is a commonly used method for long-term storage of cells, including human peripheral blood mononuclear cells (PBMCs) isolated from whole blood or leukopaks.To cryopreserve PBMCs, the cells are resuspended in cryopreservation medium, cooled to extremely low temperatures, then stored at liquid nitrogen …Objective: We investigated the effects of freeze-thawing on the properties of articular cartilage. Design: The reproducibility of repeated biomechanical assay of the same osteochondral sample was first verified with 11 patellar plugs from 3 animals. Then, 4 osteochondral samples from 15 bovine patellae were divided into 4 groups. The …The impact of cryopreservation on the sperm membrane is well known, and many studies agree that frozen–thawed semen is less fertile than fresh semen because of reduced sperm motility and a lower number of viable spermatozoa [ 27, 28 ]; however, it does not seem to negatively influence the live birth rate [ 29 ]. 2.2.Combining single-cell RNA sequencing (scRNA-seq) with upstream cell preservation procedures such as cryopreservation or methanol fixation has recently become more common. By separating cell ...Cryopreservation involves the preservation of biological materials, including cells, embryos, tissues, and organs, at ultra-low temperatures (in a state of suspended animation), ...Sep 13, 2022 · Cryopreservation is a rapidly developing field that is continually adapting to meet the challenges presented by ice and low temperatures when trying to preserve viability in larger tissues and structures. The larger structures become, the more challenging attempts at cryopreservation, using current techniques, becomes. Cryopreservation is a key enabling technology in regenerative medicine that provides stable and secure extended cell storage for primary tissue isolates and constructs and prepared cell preparations. The essential detail of the process as it can be applied to cell-based therapies is set out in this review, covering tissue and cell isolation, …Cryopreservation strives to maximize the viability and biofunctionality of cells and tissues by cooling them to a subzero temperature to facilitate storage and delivery. This technology has enabled clinics and labs to preserve rare and crucial samples and is poised to become more important with rising interest in cell therapy.toxic to cells at high concentrations, was identified as a necessary step to protect against rampant cell death during cryo-. preservation. In addition to osmotic …Cryopreservation, the process of storing materials at sub-zero temperatures, is an essential process for fundamental research, as well as the clinical, biomedical and …Cryopreservation, the process of storing materials at sub-zero temperatures, is an essential process for fundamental research, as well as the clinical, biomedical and …Cryopreservation is the method of keeping the live cells, tissues and other biological samples in a deep freeze at subzero temperatures for the storage or preservation. The …Cryopreservation. Label cryovials with the date, name of researcher, cell number, passage number and cell type (and any other useful information, for example genetic modifications). If cells are adherent, remove the cell culture media, wash in PBS, add enough trypsin to cover the cells and incubate for approximately 2 min in a 37°C incubator.Transplantation of hematopoietic stem cells (HSC) has been successfully developed as a part of treatment protocols for a large number of clinical indications, and cryopreservation of both autologous and allogeneic sources of HSC grafts is increasingly being employed to facilitate logistical challenges in coordinating the collection, processing, preparation, …Plant cryopreservation, specifically, is a process of cooling and storing vegetal structure as plant cells, tissues, or organs in liquid nitrogen (LN; −196 °C) or LN vapor (−160 °C). This methodology ensures the maintenance of samples’ viability after thawing, and indefinite storage is possible. The cryopreservation technique is based ...Cryopreservation involves storing biological material at ultra-low temperatures, usually in liquid nitrogen. This allows long-term preservation by stopping almost all metabolic activity in cells. Materials are frozen using slow freezing, rapid freezing, or stepwise freezing methods. They are then stored long-term at temperatures near -196°C.Apr 15, 2010 · Practical Applications of Plant Cryopreservation. Cryopreservation of Dedifferentiated Cells and Tissues. Traditional, Simplified, and Novel Approaches for Dedifferentiated Cell Cryopreservation. Cryopreservation of Shoots and Roots. Enhancing Survival in Cryopreserved Shoot Tips using Vitrification. Cryopreservation of Seeds and Embryos The objective of this study was to investigate the recovery of bacteria from ewe milk after freezing for 4 or 8 weeks with and without the addition of glycerol as a cryopreservant. A total of 50 udder-half milk samples with a known range of bacterial species were selected, stored and analyzed in 5 treatment-groups: time zero, frozen for 4 weeks with, and …Jun 4, 2019 · Abstract. In this concept article, we outline a variety of new approaches that have been conceived to address some of the remaining challenges for developing improved methods of biopreservation. This recognizes a true renaissance and variety of complimentary, high-potential approaches leveraging inspiration by nature, nanotechnology, the thermodynamics of pressure, and several other key fields ... Core Tip: The manuscript is an overview of current cryopreservation protocols used for cold storage of hematopoietic stem cells, mesenchymal stem cells and induced pluripotent stem cells.Although dimethyl sulfoxide (DMSO) is commonly used in cryopreservation of cell lines, primary cells and stem cells, the use of DMSO has been …Cryopreservation of tissues is a tough challenge. Cryopreservation is categorized into slow-freezing and vitrification, and vitrification has recently been recognized as a suitable method for ... As methods for cryopreservation of the tissues of origin as well as the resulting stem cell populations are optimized and reproducible using the strategies outlined, ongoing and future research in this area includes cryopreservation of three-dimensional culture systems and cell-infiltrated scaffolds. Vascularized tissues and organs require perfusion for the various stages of preservation, such as preconditioning (including pre-equilibration with CPAs), preservation, and resuscita-. Fig. 1. New nature-inspired and bioengineering approaches to cryopreservation compared with classic methods. tion/recovery. Cryopreservation is a fundamental tool of assisted reproduction and establishing long-term banking germplasm. Fertility cryopreservation is challenging because of gametes and embryos morphological, functional, and genetic changes in the cells after freeze- thawing [176,177,178,179].Abstract. Cryopreservation involves the preservation of biological materials, including cells, embryos, tissues, and organs, at ultra-low temperatures (in a state of suspended animation), for a long period of time, and in a way that allows them to be restored whenever required. Freezing of biological samples is generally accompanied by numerous ...Cryopreservation is routinely used for the long-term storage of cells in various areas of academic, industrial, and clinical research. To keep frozen cells alive, it is necessary to vitrify (or nanocrystallize) water on the inside and outside of the cells. Vitrification is conventionally achieved by adding at least one cryoprotective agent (CPA ...Cryopreservation. Label cryovials with the date, name of researcher, cell number, passage number and cell type (and any other useful information, for example genetic modifications). If cells are adherent, remove the cell culture media, wash in PBS, add enough trypsin to cover the cells and incubate for approximately 2 min in a 37°C incubator. Cryopreservation gained prominence in human medicine after its use in infertility treatment. Since then, gamete cryopreservation has been developed to combat infertility. Sperm was the first successfully frozen reproductive cell and remains the easiest to freeze due to its tiny cytoplasm and thus low water content. Dimethyl sulfoxide (DMSO) is the cryoprotectant of choice for most animal cell systems since the early history of cryopreservation. It has been used for decades in many thousands of cell transplants. …Improved cryopreservation of adipose tissue for the isolation of mesenchymal stem cells WO2010096821A2 (en) * 2009-02-23: 2010-08-26: Cell & Tissue Systems, Inc. Method for ice-free cryopreservation of tissue US20120190004A1 (en) * 2009-06-23: 2012-07-26: Biolex Therapeutics, Inc. Cryopreservation is the use of very low temperatures to preserve structurally intact living cells and tissues. Unprotected freezing is normally lethal and this chapter seeks to analyze some of the mechanisms involved and to show how cooling can be used to produce stable conditions that preserve life. The biological effects of cooling are ... Aug 15, 2020 · Cryopreservation and freeze-drying can be used to preserve cells or tissues for prolonged periods. Vitrification, or ice-free cryopreservation, is an alternative to cryopreservation that enables cooling cells to cryogenic temperatures in the absence of ice. The processing pathways involved in (ice-free) cryopreservation and freeze-drying of ... Cryoprotectants are basically some chemical compounds which prevent cells or tissues from damage due to freezing. Mostly vitrification and thawing process are …Core Tip: The manuscript is an overview of current cryopreservation protocols used for cold storage of hematopoietic stem cells, mesenchymal stem cells and induced pluripotent stem cells.Although dimethyl sulfoxide (DMSO) is commonly used in cryopreservation of cell lines, primary cells and stem cells, the use of DMSO has been … In addition to general considerations, cell-specific recommendations for hepatocytes, pancreatic islets, sperm, oocytes, and stem cells should be observed to maximize yields. For example, rapid cooling is associated with better cryopreservation outcomes for oocytes, pancreatic islets, and embryonic stem cells while slow cooling is recommended ... Cryopreservation, the process of storing materials at sub-zero temperatures, is an essential process for fundamental research, as well as the clinical, biomedical and food sciences 1. The ability to prolong the storage of biological materials, by reducing the temperature to slow the rate of degradation, has wide-reaching applications.Apr 1, 2021 · Cryopreservation of testicular cells and tissues is useful for the preservation and restoration of fertility in pre-pubertal males expecting gonadotoxic treatment for cancer and genetic diseases causing impaired spermatogenesis. A number of freezing and vitrification protocols have thus been tried and variable results have been reported in terms of cell viability spermatogenesis progression ... Cryopreservation using DMSO resulted in high percentages of activated platelets; namely 54% of the recovered 94%. When using trehalose, 98% of the platelets had intact membranes after freezing and thawing, whereas 76% were not activated. Using Fourier transform infrared spectroscopy, subzero membrane phase behavior of platelets has …Plant cryopreservation is useful for long term storage of clonal germplasm and endangered species. Clonally propagated crops which produce recalcitrant seeds cannot be easily conserved using conventional methods. Preservation of plants in vitro is limited to two years and not ideal for germplasm storage for a very long time. The need …Cryopreservation involves the preservation of biological materials, including cells, embryos, tissues, and organs, at ultra-low temperatures (in a state of suspended animation), ...Examples of how to use “cryopreservation” in a sentence from Cambridge Dictionary.There are plenty of practical reasons not to sign up for whole body cryopreservation after death - the most important of which is that there is absolutely no proof or guarantee that it is reversible. There is a significant and widespread need to extend the applicability of long-term cold storage to a wider range of insect species, life stages, cells, and tissues. Practical applications for cryopreservation are abundant among the many disciplines utilizing insects or their cells. In most cases when whole insects are currently subjected to ... Cryopreservation is a critical procedure in autologous hematopoietic stem cell transplantation. Dimethyl sulfoxide (DMSO) is the cryoprotectant of choice. Optimization of the cryopreservation protocol in the past revealed a dramatic loss of cell viability associated with a reduction of the DMSO concentration below 2 vol % in the freezing … Vascularized tissues and organs require perfusion for the various stages of preservation, such as preconditioning (including pre-equilibration with CPAs), preservation, and resuscita-. Fig. 1. New nature-inspired and bioengineering approaches to cryopreservation compared with classic methods. tion/recovery. Wipe ampoule with a tissue soaked in 70% alcohol prior to opening. Pipette the whole content of the ampoule into a sterile tube (e.g., 15 mL capacity). Then slowly add 5 mL pre-warmed medium that has already been supplemented with the appropriate constituents. Determine the viable cell density using trypan blue.Jul 18, 2022 · Cryopreservation, the process of storing materials at sub-zero temperatures, is an essential process for fundamental research, as well as the clinical, biomedical and food sciences 1. The ability... Vitrification—glass formation without crystallization [ 6, 7 ]—is one of the basic principles used in plant cryopreservation methods. Only a few plants can form a vitreous state naturally [ 8 ]. Most plants cannot vitrify themselves because they lack glassy state inducing substances and/or have high water content.Feb 1, 2021 · 1 Introduction. Cryopreservation is a basic and important technique to achieve long-term storage of organs, tissues, cells, and other biological materials by using a very low temperature (at −80 or −196 °C). The addition of non-permeating CPAs sugars in cryopreservation medium is one of the approaches to overcome the problems limiting the success of cell viability after warming . Notably, different saccharides in non-permeating CPAs can have different protective behavior, due to their physical and chemical properties [ 120 ] .
There is a significant and widespread need to extend the applicability of long-term cold storage to a wider range of insect species, life stages, cells, and tissues. Practical applications for cryopreservation are abundant among the many disciplines utilizing insects or their cells. In most cases when whole insects are currently subjected to ... . Turtle beach barbados location
Nov 16, 2022 · Cryopreservation is the process of pres erving living cells and tissues at very low temperatures for an. extended duration of time [8]. Types of cryopreservation processes can typically be ... Cryopreservation can be an efficient method of maintaining the potential of raising live mice of the thousands of genetically modified genotypes currently available ( Songsasen and Leibo, 1998 ). It can serve as a fail-safe measure, should a strain become genetically contaminated. In addition to being used for murine reproductive purposes ...Oocyte cryopreservation is a reliable method for the maintenance of fertility in women undergoing treatment for medical conditions such as cancer or …While cryopreservation strategies for specific cells (such as red blood cells) are successful and widely implemented, preservation of other cell types, tissues and whole organs remains challenging. This is due to the multifactorial nature of freeze-thaw damage, the complexity of preserving biological matter and the (country-to-country) variability of …Background. Cryopreservation of human ovarian tissue has been increasingly applied worldwide to safeguard fertility in cancer patients, notably in young girls and women who cannot delay the onset of their treatment. Moreover, it has been proposed to patients with benign pathologies with a risk of premature ovarian insufficiency.While stem cell cryopreservation methods have been optimized using dimethylsulfoxide (DMSO), the established techniques are not optimal when applied to unfertilized human embryonic cells. In addition, important questions remain regarding the toxicity and characteristics of DMSO for treatment of stem cells for clinical use. The …The addition of non-permeating CPAs sugars in cryopreservation medium is one of the approaches to overcome the problems limiting the success of cell viability after warming . Notably, different saccharides in non-permeating CPAs can have different protective behavior, due to their physical and chemical properties [ 120 ] .Embryo Technologies in South American Camelids. MARCELO H. RATTO, GREGG P. ADAMS, in Current Therapy in Large Animal Theriogenology (Second Edition), 2007 Embryo Cryopreservation. The effects of two cryoprotectants, propylene glycol and ethylene glycol, on post-thaw reexpansion and morphology of blastocysts have been …Jun 9, 2022 · Cryopreservation is performed either through programmable slow freezing, vitrification or low-CPA vitrification (ultra-rapid cooling) with vitrification being is the preferred technique in cryonics because it prevents/reduces the formation of damaging ice crystals within the cryopreserved subject . Dec 13, 2021 · Plant cryopreservation, specifically, is a process of cooling and storing vegetal structure as plant cells, tissues, or organs in liquid nitrogen (LN; −196 °C) or LN vapor (−160 °C). This methodology ensures the maintenance of samples’ viability after thawing, and indefinite storage is possible. The cryopreservation technique is based ... Cryoprotectants play a vital role in the cryopreservation process, protecting biological samples from freezing damage. Here, we evaluate the effects of the combination and interaction of different extenders with permeable and non-permeable cryoprotectants, on the cryopreservation of Danio rerio sperm, analyzing the effects of cryopreservation …Abstract. Cryopreservation involves the preservation of biological materials, including cells, embryos, tissues, and organs, at ultra-low temperatures (in a state of suspended animation), for a long period of time, and in a way that allows them to be restored whenever required. Freezing of biological samples is generally accompanied by numerous ...The addition of non-permeating CPAs sugars in cryopreservation medium is one of the approaches to overcome the problems limiting the success of cell viability after warming . Notably, different saccharides in non-permeating CPAs can have different protective behavior, due to their physical and chemical properties [ 120 ] .Cryopreservation by control cooling rate is an expensive method only viable for specific cases such as prokaryote organisms 3-5. When an aqueous solution is rapidly cooled, crystallization is avoided resulting in water being in a metastable amorphous solid before becoming vitrified in a vitreous solid at temperatures below the glass transition.Objective: We investigated the effects of freeze-thawing on the properties of articular cartilage. Design: The reproducibility of repeated biomechanical assay of the same osteochondral sample was first verified with 11 patellar plugs from 3 animals. Then, 4 osteochondral samples from 15 bovine patellae were divided into 4 groups. The …May 25, 2023 · Cryopreservation involves maintaining the physiological function of cells, tissues, and organs at ultra-low temperatures (−80°C or −196°C), at which point cellular activities nearly cease. Cryopreservation can effectively halt biological time and overcome the limitations of existing low-temperature storage technologies, making it a ... May 27, 2021 · Traditional cryopreservation is a highly effective method for storing cells and tissues, which works by keeping cells in “suspended animation”, as Sameena Iqbal, bioresource manager at the Wellcome Sanger Institute explains. “By freezing cells, it stops the metabolic activity and preserves the compounds within cells such as enzymes.”. Dimethyl sulfoxide (DMSO) is used as a cryopreservant for mammalian cell cultures because . it is an organic solvent. it easily penetrates cells. it protects cells by preventing crystallization of water. it is also utilized as a nutrient .Cryopreservation is a process that preserves organelles, cells, tissues, or any other biological constructs by cooling the samples to very low temperatures. The responses of living cells to ice formation are of theoretical interest and practical relevance. Stem cells and other viable tissues, which …Cryopreservation - Download as a PDF or view online for free. 2. Introduction Cryopreservation is the technique of freezing cells and tissues at very low temperatures (sub-zero temperatures, typically -196oC) at which the biological material remains genetically stable and metabolically inert, while minimizing ice crystal formation. ….