The effect of local cold injury in rats on the osmotic resistance of erythrocytes in the early and late reactive terms was investigated. Cryoinfluence was performed by pressing the applicator to the skin on the lateral surface of the rat thigh for 30, 60 and 120 s. Afterwards, the blood was collected from the aorta of animals after 1 and 4 hours and after 24 and 48 hours (early and late reactive periods, respectively). The effect of hypotonic solutions on cells was used as a test for osmotic resistance of erythrocytes. It is shown that the osmotic resistance of erythrocytes in the early reactive term increases (compared with control cells). In the late reactive term, the indices of osmotic resistance of erythrocytes after 24 hours approached the control values and after 48 hours they reached the norm. Osmotic resistance of erythrocytes in both reactive terms after local cold injury does not depend on the duration of cryoinfluence (30, 60, 120 s). The findings are considered from the standpoint of adaptive reactions that have an allostatic nature.
{"title":"Osmotic Resistance of Rat’s Erythrocytes After Local Cold Injury","authors":"G. Kovalov, O. Chabanenko, N. Orlova, N. Shpakova","doi":"10.15407/cryo32.01.024","DOIUrl":"https://doi.org/10.15407/cryo32.01.024","url":null,"abstract":"The effect of local cold injury in rats on the osmotic resistance of erythrocytes in the early and late reactive terms was investigated. Cryoinfluence was performed by pressing the applicator to the skin on the lateral surface of the rat thigh for 30, 60 and 120 s. Afterwards, the blood was collected from the aorta of animals after 1 and 4 hours and after 24 and 48 hours (early and late reactive periods, respectively). The effect of hypotonic solutions on cells was used as a test for osmotic resistance of erythrocytes. It is shown that the osmotic resistance of erythrocytes in the early reactive term increases (compared with control cells). In the late reactive term, the indices of osmotic resistance of erythrocytes after 24 hours approached the control values and after 48 hours they reached the norm. Osmotic resistance of erythrocytes in both reactive terms after local cold injury does not depend on the duration of cryoinfluence (30, 60, 120 s). The findings are considered from the standpoint of adaptive reactions that have an allostatic nature.","PeriodicalId":53457,"journal":{"name":"Problems of Cryobiology and Cryomedicine","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2022-03-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"46205370","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
V. Lebedinets, L. Ostankova, M. Bondarovych, O. Lutsenko, D. Lebedinets, Igor Grisha, M. Ostankov, K. Parkhomenko, A. Goltsev
The paper outlines the experimental data on the impact of lyophilization on preservation of structural and functional characteristics of cells from human cord blood leukoconcentrate (HCBL). Lyophilization of HCBL was shown to ensure the preservation of cells with immune modulating potential in a heterogeneous population of HCBL. Lyophilized HCBL (lHCBL), to the same extent as cryopreserved one (cHCBL), revealed an equal immune correcting effect during ischemic brain injury in vivo in the experimental model of ischemic stroke (IS). The inclusion of lHCBL in therapy of brain ischemia demonstrated the improved indices of IFN-ү+ and IL-10+ spleen cells, adhesive and phagocytic activity of peritoneal cavity cells in rats with IS. The efficiency of using both lHCBL and cHCBL during IS may be associated with the implementation of therapeutic effect by presented among them cells and mediators, including multi-vector regulatory systems that maintain homeostatic stability of the body (immune, endocrine, nervous ones).
{"title":"Lyophilized Human Cord Blood Leukoconcentrate to Treat Brain Ischemia in Rats","authors":"V. Lebedinets, L. Ostankova, M. Bondarovych, O. Lutsenko, D. Lebedinets, Igor Grisha, M. Ostankov, K. Parkhomenko, A. Goltsev","doi":"10.15407/cryo32.01.044","DOIUrl":"https://doi.org/10.15407/cryo32.01.044","url":null,"abstract":"The paper outlines the experimental data on the impact of lyophilization on preservation of structural and functional characteristics of cells from human cord blood leukoconcentrate (HCBL). Lyophilization of HCBL was shown to ensure the preservation of cells with immune modulating potential in a heterogeneous population of HCBL. Lyophilized HCBL (lHCBL), to the same extent as cryopreserved one (cHCBL), revealed an equal immune correcting effect during ischemic brain injury in vivo in the experimental model of ischemic stroke (IS). The inclusion of lHCBL in therapy of brain ischemia demonstrated the improved indices of IFN-ү+ and IL-10+ spleen cells, adhesive and phagocytic activity of peritoneal cavity cells in rats with IS. The efficiency of using both lHCBL and cHCBL during IS may be associated with the implementation of therapeutic effect by presented among them cells and mediators, including multi-vector regulatory systems that maintain homeostatic stability of the body (immune, endocrine, nervous ones).","PeriodicalId":53457,"journal":{"name":"Problems of Cryobiology and Cryomedicine","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2022-03-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"46012625","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
The history of the development and use of low-temperature electron microscopic methods of freeze-fracture, freeze-substitution and others in cryobiological research at the Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine is presented in this article. These methods’ possibilities in studying the processes of crystal formation in cryoprotectant solutions, cell suspensions, and tissues under various freezing conditions are demonstrated. Some results of the analysis of ultrastructural changes in biological systems of various organization levels, at different stages of ontogenesis, under the influence of cooling in a wide temperature range (from 37 to –196°C) are presented. The use of a high resolution electron microscopic method in combination with an accessory technical equipment and some methodological techniques allowed to obtain fundamental results important for cryobiology on ice crystals formation and localization in the intracellular volume, the temperature-dependent transmembrane proteins redistribution, changes in the ultrastructure of erythrocytes and their membranes during hypothermic storage.
{"title":"Opportunities of Electron Microscopy When Solving Cryobiological Tasks. Retrospective Analysis","authors":"N. Repin, L. Marchenko, T. Govorukha, A. Goltsev","doi":"10.15407/cryo32.01.003","DOIUrl":"https://doi.org/10.15407/cryo32.01.003","url":null,"abstract":"The history of the development and use of low-temperature electron microscopic methods of freeze-fracture, freeze-substitution and others in cryobiological research at the Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine is presented in this article. These methods’ possibilities in studying the processes of crystal formation in cryoprotectant solutions, cell suspensions, and tissues under various freezing conditions are demonstrated. Some results of the analysis of ultrastructural changes in biological systems of various organization levels, at different stages of ontogenesis, under the influence of cooling in a wide temperature range (from 37 to –196°C) are presented. The use of a high resolution electron microscopic method in combination with an accessory technical equipment and some methodological techniques allowed to obtain fundamental results important for cryobiology on ice crystals formation and localization in the intracellular volume, the temperature-dependent transmembrane proteins redistribution, changes in the ultrastructure of erythrocytes and their membranes during hypothermic storage.","PeriodicalId":53457,"journal":{"name":"Problems of Cryobiology and Cryomedicine","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2022-03-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"47602241","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
A. Zavhorodnii, V. Bilushko, A. Paliy, M. Kalashnyk, N. Kalashnyk, A. Paliy
{"title":"Freeze-Drying Improved the Stability of Tuberculin Purified Protein Derivative (PPD) in Mammals","authors":"A. Zavhorodnii, V. Bilushko, A. Paliy, M. Kalashnyk, N. Kalashnyk, A. Paliy","doi":"10.15407/cryo32.01.063","DOIUrl":"https://doi.org/10.15407/cryo32.01.063","url":null,"abstract":"","PeriodicalId":53457,"journal":{"name":"Problems of Cryobiology and Cryomedicine","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2022-03-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"48614167","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Sara Campos, E. Paredes, Jesús Troncoso Jesús Troncoso
{"title":"Can Sea Urchin Eggs Shelf-Life Be Extended by Cold Storage?","authors":"Sara Campos, E. Paredes, Jesús Troncoso Jesús Troncoso","doi":"10.15407/cryo32.01.068","DOIUrl":"https://doi.org/10.15407/cryo32.01.068","url":null,"abstract":"","PeriodicalId":53457,"journal":{"name":"Problems of Cryobiology and Cryomedicine","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2022-03-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"49080771","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
The effect of synthetic leu-enkephalin (dalargin) on apoptosis and necrosis in leukocytes after cold stress on the morphological characteristics of cells and using fluorescent dyes Hoechst 33342 as well as propidium iodide was studied. The degree of cold stress influence depending on the duration of incubation of cell suspension at 0–2 and 37°C on the initiation of apoptosis and necrosis on the morphological characteristics of cells was determined. It has been proven that the ratio of apoptotic cells to necrotic ones was maximal after the use of the regimen as follows: preliminary incubation of leukocytes at 37°C – 15 min, cell exposure at 0–2°C – 15 min and subsequent incubation of leukocytes at 37°C. It has been confirmed that pre-incubation of cells with neuropeptide to cold stress affects the effectiveness of the protective action of dalargin, in particular, contributes to a significant (p ≤ 0.05) reduction in the percentage of leukocytes with morphological signs of apoptosis and necrosis. It was found that a consistent increase in temperature after cold stress to 37°C did not affect the percentage of cells with morphological signs of apoptosis and necrosis. It has been validated that the percentage of leukocytes with the apoptosis and necrosis signs after cold stress was significantly reduced after incubation of leukocytes with dalargin at a concentration of 10–9 mol/l.
{"title":"Effect of Leu-Enkephalin (Dalargin) on Apoptosis and Necrosis of Leukocytes After Cold Stress","authors":"N. Moisieieva, O. Gulevskyy, O. Gorina","doi":"10.15407/cryo32.01.014","DOIUrl":"https://doi.org/10.15407/cryo32.01.014","url":null,"abstract":"The effect of synthetic leu-enkephalin (dalargin) on apoptosis and necrosis in leukocytes after cold stress on the morphological characteristics of cells and using fluorescent dyes Hoechst 33342 as well as propidium iodide was studied. The degree of cold stress influence depending on the duration of incubation of cell suspension at 0–2 and 37°C on the initiation of apoptosis and necrosis on the morphological characteristics of cells was determined. It has been proven that the ratio of apoptotic cells to necrotic ones was maximal after the use of the regimen as follows: preliminary incubation of leukocytes at 37°C – 15 min, cell exposure at 0–2°C – 15 min and subsequent incubation of leukocytes at 37°C. It has been confirmed that pre-incubation of cells with neuropeptide to cold stress affects the effectiveness of the protective action of dalargin, in particular, contributes to a significant (p ≤ 0.05) reduction in the percentage of leukocytes with morphological signs of apoptosis and necrosis. It was found that a consistent increase in temperature after cold stress to 37°C did not affect the percentage of cells with morphological signs of apoptosis and necrosis. It has been validated that the percentage of leukocytes with the apoptosis and necrosis signs after cold stress was significantly reduced after incubation of leukocytes with dalargin at a concentration of 10–9 mol/l.","PeriodicalId":53457,"journal":{"name":"Problems of Cryobiology and Cryomedicine","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2022-03-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"43818219","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
A. Goltsev, K. Yampolska, H. Kisielova, Maksym Оstankov, T. Dubrava, N. Babenko, Yuliya Gaevska, M. Bondarovych
An important component of the pathogenesis of autoimmune diseases is the immune system deregulation as an impaired tolerance to its own antigens by reducing the content of T-regulatory cells. Their formation is closely related to the function of dendritic cells (DCs), so in autoimmune diseases the use of DCs with tolerogenic potential is promising for the restoration of antigen-specific tolerance. Recently, the issue of establishing the banks of tolerogenic DCs for clinical use, which involves their cryopreservation, has been actively discussed. To date, there is no common protocol for DCs freezing, which would take into account the different sources of their obtaining, the initial structural and functional state before freezing, composition of cryopreservation media and other factors. The review summarizes experimental data on cryopreservation of peripheral blood and bone marrow mononuclear cells or monocytes. The potential for their further ex vivo differentiation into DCs to ensure the stability of immature phenotype and tolerogenic function has been studied.
{"title":"Cryopreservation as Biotechnological Application of Dendritic Cells in Clinical Practice","authors":"A. Goltsev, K. Yampolska, H. Kisielova, Maksym Оstankov, T. Dubrava, N. Babenko, Yuliya Gaevska, M. Bondarovych","doi":"10.15407/cryo31.04.289","DOIUrl":"https://doi.org/10.15407/cryo31.04.289","url":null,"abstract":"An important component of the pathogenesis of autoimmune diseases is the immune system deregulation as an impaired tolerance to its own antigens by reducing the content of T-regulatory cells. Their formation is closely related to the function of dendritic cells (DCs), so in autoimmune diseases the use of DCs with tolerogenic potential is promising for the restoration of antigen-specific tolerance. Recently, the issue of establishing the banks of tolerogenic DCs for clinical use, which involves their cryopreservation, has been actively discussed. To date, there is no common protocol for DCs freezing, which would take into account the different sources of their obtaining, the initial structural and functional state before freezing, composition of cryopreservation media and other factors. The review summarizes experimental data on cryopreservation of peripheral blood and bone marrow mononuclear cells or monocytes. The potential for their further ex vivo differentiation into DCs to ensure the stability of immature phenotype and tolerogenic function has been studied.","PeriodicalId":53457,"journal":{"name":"Problems of Cryobiology and Cryomedicine","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2021-12-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"49305403","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Оlha Denysova, G. Zhegunov, T. Yakimenko, N. Hladka, V. Prichodchenko, Olha Bobrutska
Babesiosis is one of the most common protozoan diseases in dogs, which in the absence of the necessary veterinary care often leads to the animal death. The changes of hematological parameters in dogs during complex treatment of babesiosis and transfusion of cryopreserved erythrocytes were evaluated in the research. Erythrocytes were frozen using 20% DMSO by immersion into liquid nitrogen. Transfusion of cryopreserved erythrocytes together with antiparasitic chemotherapy has been shown to promote rapid recovery of hematological parameters of animal blood (hemoglobin, hematocrit and erythrocyte counts), as well as improves their well-being, promotes faster recovery. Cryopreserved with DMSO erythrocytes can be recommended for long-term storage of blood at cryobanks.
{"title":"Blood Characteristics in Dogs During Treatment of Babesiosis Using Transfusion of Cryopreserved Erythrocytes","authors":"Оlha Denysova, G. Zhegunov, T. Yakimenko, N. Hladka, V. Prichodchenko, Olha Bobrutska","doi":"10.15407/cryo31.04.304","DOIUrl":"https://doi.org/10.15407/cryo31.04.304","url":null,"abstract":"Babesiosis is one of the most common protozoan diseases in dogs, which in the absence of the necessary veterinary care often leads to the animal death. The changes of hematological parameters in dogs during complex treatment of babesiosis and transfusion of cryopreserved erythrocytes were evaluated in the research. Erythrocytes were frozen using 20% DMSO by immersion into liquid nitrogen. Transfusion of cryopreserved erythrocytes together with antiparasitic chemotherapy has been shown to promote rapid recovery of hematological parameters of animal blood (hemoglobin, hematocrit and erythrocyte counts), as well as improves their well-being, promotes faster recovery. Cryopreserved with DMSO erythrocytes can be recommended for long-term storage of blood at cryobanks.","PeriodicalId":53457,"journal":{"name":"Problems of Cryobiology and Cryomedicine","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2021-12-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"46072444","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
V. Kiroshka, Yu. O. Bozhkova, A. Gavas, I. Rula, T. Bondarenko, G. Semko, V. Pyatikop
Morphological characteristics and function of ovarian tissue after 2–120 hrs of storage at 4 and 37°C in incubation media with different composition (mannitol-containing solution (MСS) and phosphate buffered saline (PBS)) were investigated. The function of ovarian tissue after normo- and hypothermic storage was studied by heterotopic transplantation to recipient animals simultaneously with bilateral ovariectomy. After 2-hour storage at 37ºC and 24-hour one at 4ºC in all the studied media, the estradiol and progesterone levels in plasma of recipient animals were experimentally proved to be 18.2–27.5 and 13.5–18.5 ng/ml, respectively, (after fresh tissue transplantation these values for estradiol and progesterone made (28.1 ± 3.6) pg/ml and (19.1 ± 2.8) ng/ml, respectively). Prolongation of tissue storage time at 37ºC increased the number of degenerative follicles, augmented TBARS concentration and reduced sex hormone levels after transplantation using both MCS and PBS. A protective effect of MCS only, manifested in endocrine function restoration in recipient animals after transplantation ((13.5 ± 2.5) pg/ml of estradiol) was found after 48 hrs of ovarian tissue hypothermic storage. If PBS was used, the estradiol level was within the indices for ovariectomized animals ((7.1 ± 1.5 (pg/ml)).
{"title":"Endocrine Function of Ovarian Tissue Grafts After Normo- and Hypothermic Storage","authors":"V. Kiroshka, Yu. O. Bozhkova, A. Gavas, I. Rula, T. Bondarenko, G. Semko, V. Pyatikop","doi":"10.15407/cryo31.04.326","DOIUrl":"https://doi.org/10.15407/cryo31.04.326","url":null,"abstract":"Morphological characteristics and function of ovarian tissue after 2–120 hrs of storage at 4 and 37°C in incubation media with different composition (mannitol-containing solution (MСS) and phosphate buffered saline (PBS)) were investigated. The function of ovarian tissue after normo- and hypothermic storage was studied by heterotopic transplantation to recipient animals simultaneously with bilateral ovariectomy. After 2-hour storage at 37ºC and 24-hour one at 4ºC in all the studied media, the estradiol and progesterone levels in plasma of recipient animals were experimentally proved to be 18.2–27.5 and 13.5–18.5 ng/ml, respectively, (after fresh tissue transplantation these values for estradiol and progesterone made (28.1 ± 3.6) pg/ml and (19.1 ± 2.8) ng/ml, respectively). Prolongation of tissue storage time at 37ºC increased the number of degenerative follicles, augmented TBARS concentration and reduced sex hormone levels after transplantation using both MCS and PBS. A protective effect of MCS only, manifested in endocrine function restoration in recipient animals after transplantation ((13.5 ± 2.5) pg/ml of estradiol) was found after 48 hrs of ovarian tissue hypothermic storage. If PBS was used, the estradiol level was within the indices for ovariectomized animals ((7.1 ± 1.5 (pg/ml)).","PeriodicalId":53457,"journal":{"name":"Problems of Cryobiology and Cryomedicine","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2021-12-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"49355585","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
I. Vysekantsev, V. Martsenyuk, I. Buriak, T. Gurina
The effect of DMSO and sucrose added to alginate gel on the viability of immobilized Saccharomyces boulardii yeast cells after rapid and slow cooling to –196 °C has been studied. Thermomechanical analysis revealed the inflection of phase transitions corresponding to each component of the solutions when cooling those of 1% sodium alginate contained 20% sucrose or 5% DMSO down to –196 °C. Phase transitions caused by the presence of DMSO and sucrose in sodium alginate solutions were observed at lower temperatures than in impurity-free sodium alginate solution. In experiments to study the viability of free and immobilized yeast cells after freezing, it was found that the highest viability of free cells suspended in solutions of sodium alginate, DMSO, sucrose, and the cells immobilized in gel granules, was provided by a cooling rate of 1 deg/min followed by immersion in liquid nitrogen. After adding DMSO and sucrose separately to the alginate gel, as well as their combinations, the viability of immobilized cells increased. The maximum viability of immobilized cells was found in gel granules, which contained the following combinations of cryoprotectants: 5% DMSO and 10% sucrose, 5% DMSO and 20% sucrose, 10% DMSO and 10% sucrose and 10% DMSO and 20% sucrose.
{"title":"Freezing Regimens and Gel Carrier Composition Influence Safety of Saccharomyces boulardii Immobilized Yeast Cells","authors":"I. Vysekantsev, V. Martsenyuk, I. Buriak, T. Gurina","doi":"10.15407/cryo31.04.343","DOIUrl":"https://doi.org/10.15407/cryo31.04.343","url":null,"abstract":"The effect of DMSO and sucrose added to alginate gel on the viability of immobilized Saccharomyces boulardii yeast cells after rapid and slow cooling to –196 °C has been studied. Thermomechanical analysis revealed the inflection of phase transitions corresponding to each component of the solutions when cooling those of 1% sodium alginate contained 20% sucrose or 5% DMSO down to –196 °C. Phase transitions caused by the presence of DMSO and sucrose in sodium alginate solutions were observed at lower temperatures than in impurity-free sodium alginate solution. In experiments to study the viability of free and immobilized yeast cells after freezing, it was found that the highest viability of free cells suspended in solutions of sodium alginate, DMSO, sucrose, and the cells immobilized in gel granules, was provided by a cooling rate of 1 deg/min followed by immersion in liquid nitrogen. After adding DMSO and sucrose separately to the alginate gel, as well as their combinations, the viability of immobilized cells increased. The maximum viability of immobilized cells was found in gel granules, which contained the following combinations of cryoprotectants: 5% DMSO and 10% sucrose, 5% DMSO and 20% sucrose, 10% DMSO and 10% sucrose and 10% DMSO and 20% sucrose.","PeriodicalId":53457,"journal":{"name":"Problems of Cryobiology and Cryomedicine","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2021-12-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"48924808","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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