The present study was conducted with the goal of evaluating the viability of spermatozoa of the Deccan mahseer (Tor khudree, Cyprinidae) cryopreserved using different strategies. Immotile spermatozoa pooled from 2–4 males were diluted with modified fish Ringer's solution (pH: 7.48) and protected with dimethyl sulfoxide (Me2SO) at 5%, 10%, and 15% and subjected to different equilibration periods. Diluted samples (1:10) were drawn into 500 µL plastic straws and frozen at a distance of 5 cm from the level of liquid nitrogen (LN2) and preserved for 385 days in LN2. Me2SO at 10% resulted in higher post-thaw spermatozoa motility rate (46.7%) than 5% or 15% (up to 33.3%) after 385 days of cryopreservation. Of the different equilibration periods, 20–40 min generally produced higher motility (%) rates than 0, 10, 50, 60, 70, 80, or 90 min. The highest post-thaw motility of spermatozoa was obtained when they were frozen at 2 cm (−120.3°C) above the level of LN2 and the optimum freezing rate was found to be 14.5 ± 0...
{"title":"Cryopreservation of the Endangered Mahseer (Tor khudree) Spermatozoa: Effect of Dimethyl Sulfoxide, Freezing, Activating Media, and Cryostorage on Post-Thaw Spermatozoa Motility and Fertility","authors":"N. Basavaraja, S. N. Hegde, K. Palaksha","doi":"10.1089/CPT.2006.4.31","DOIUrl":"https://doi.org/10.1089/CPT.2006.4.31","url":null,"abstract":"The present study was conducted with the goal of evaluating the viability of spermatozoa of the Deccan mahseer (Tor khudree, Cyprinidae) cryopreserved using different strategies. Immotile spermatozoa pooled from 2–4 males were diluted with modified fish Ringer's solution (pH: 7.48) and protected with dimethyl sulfoxide (Me2SO) at 5%, 10%, and 15% and subjected to different equilibration periods. Diluted samples (1:10) were drawn into 500 µL plastic straws and frozen at a distance of 5 cm from the level of liquid nitrogen (LN2) and preserved for 385 days in LN2. Me2SO at 10% resulted in higher post-thaw spermatozoa motility rate (46.7%) than 5% or 15% (up to 33.3%) after 385 days of cryopreservation. Of the different equilibration periods, 20–40 min generally produced higher motility (%) rates than 0, 10, 50, 60, 70, 80, or 90 min. The highest post-thaw motility of spermatozoa was obtained when they were frozen at 2 cm (−120.3°C) above the level of LN2 and the optimum freezing rate was found to be 14.5 ± 0...","PeriodicalId":51233,"journal":{"name":"Cell Preservation Technology","volume":"138 1","pages":"31-45"},"PeriodicalIF":0.0,"publicationDate":"2006-04-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/CPT.2006.4.31","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"60913297","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}
C. Oliveri, M. Frequin, G. Malferrari, Giuliana Saltini, M. Gramegna, R. Tagliabue, P. Blasio, I. Biunno, L. Biagiotti
Several procedures to isolate DNA from difficult sources have previously been described, but they are often expensive, time consuming, and have limited applications. In this paper we describe a simple and versatile protocol to isolate nucleic acids from different plant tissues, using a silica-based extraction method. This extraction process efficiently purifies DNA from several plant cells. The obtained DNA has successfully been applied in polymerase chain reaction (PCR) assays for OGM purposes. The protocol is compatible with at least two automated liquid handling systems making it suitable for large-scale screening applications.
{"title":"A Simple Extraction Method Useful to Purify DNA from Difficult Biologic Sources","authors":"C. Oliveri, M. Frequin, G. Malferrari, Giuliana Saltini, M. Gramegna, R. Tagliabue, P. Blasio, I. Biunno, L. Biagiotti","doi":"10.1089/CPT.2006.4.51","DOIUrl":"https://doi.org/10.1089/CPT.2006.4.51","url":null,"abstract":"Several procedures to isolate DNA from difficult sources have previously been described, but they are often expensive, time consuming, and have limited applications. In this paper we describe a simple and versatile protocol to isolate nucleic acids from different plant tissues, using a silica-based extraction method. This extraction process efficiently purifies DNA from several plant cells. The obtained DNA has successfully been applied in polymerase chain reaction (PCR) assays for OGM purposes. The protocol is compatible with at least two automated liquid handling systems making it suitable for large-scale screening applications.","PeriodicalId":51233,"journal":{"name":"Cell Preservation Technology","volume":"4 1","pages":"51-54"},"PeriodicalIF":0.0,"publicationDate":"2006-04-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/CPT.2006.4.51","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"60913820","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}
With the increased recognition that cord blood is a viable source of stem cells that can be used successfully for the treatment of blood and genetic disorders, cord blood banks are faced with the challenge of developing large inventories of diverse human leukocyte antigen (HLA) phenotypes. Efficient umbilical cord blood banking requires adequate systems to reduce the volume of the cord blood unit for storage without nucleated cell or progenitor cell loss, without contamination, and with minimal risks of processing errors. Increasing regulation within cord blood banking leads to safer products, while at the same time requires standardized, reliable processes. This paper examines hydroxyethyl starch, the bottom and top, and the automated cell processing methods currently in use worldwide, and future developments bringing increased automation to the volume reduction of cord blood donations.
{"title":"Cord Blood Processing: Volume Reduction","authors":"S. Armitage","doi":"10.1089/CPT.2006.4.9","DOIUrl":"https://doi.org/10.1089/CPT.2006.4.9","url":null,"abstract":"With the increased recognition that cord blood is a viable source of stem cells that can be used successfully for the treatment of blood and genetic disorders, cord blood banks are faced with the challenge of developing large inventories of diverse human leukocyte antigen (HLA) phenotypes. Efficient umbilical cord blood banking requires adequate systems to reduce the volume of the cord blood unit for storage without nucleated cell or progenitor cell loss, without contamination, and with minimal risks of processing errors. Increasing regulation within cord blood banking leads to safer products, while at the same time requires standardized, reliable processes. This paper examines hydroxyethyl starch, the bottom and top, and the automated cell processing methods currently in use worldwide, and future developments bringing increased automation to the volume reduction of cord blood donations.","PeriodicalId":51233,"journal":{"name":"Cell Preservation Technology","volume":"4 1","pages":"9-16"},"PeriodicalIF":0.0,"publicationDate":"2006-04-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/CPT.2006.4.9","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"60914390","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}
J. Gutman, E. Shpall, Y. Nieto, P. Mcsweeney, I. Mcniece
Ex vivo expanded peripheral blood progenitor cells (PBPC) have been shown to provide rapid neutrophil engraftment, and in some patients, to eliminate neutropenia after transplantation to support high-dose chemotherapy. However, the effect of expansion culture on stem cell content and potential loss of stem cells caused by induction of differentiation remains a concern. We have transplanted 21 patients with breast cancer with expanded autologous PBPC, with 11 patients receiving expanded PBPC as their sole hematopoietic cell source. In these studies, the CD34+ cells were selected and cultured for 10 days in defined media containing 100 ng/mL each of recombinant human stem cell factor (rhSCF), recombinant human granulocyte colony stimulating factor (rhG-CSF), and recombinant human megakaryocyte growth and developmental factor (rhMGDF) in 1-liter Teflon bags at 20,000 to 50,000 cells/mL. After culture the cells were washed and reinfused after high-dose chemotherapy followed by daily administration of rhG-CSF....
{"title":"Long-Term Follow-Up of Patients with Breast Cancer Transplanted with Autologous Ex Vivo Expanded Peripheral Blood Progenitor Cells","authors":"J. Gutman, E. Shpall, Y. Nieto, P. Mcsweeney, I. Mcniece","doi":"10.1089/CPT.2006.4.4","DOIUrl":"https://doi.org/10.1089/CPT.2006.4.4","url":null,"abstract":"Ex vivo expanded peripheral blood progenitor cells (PBPC) have been shown to provide rapid neutrophil engraftment, and in some patients, to eliminate neutropenia after transplantation to support high-dose chemotherapy. However, the effect of expansion culture on stem cell content and potential loss of stem cells caused by induction of differentiation remains a concern. We have transplanted 21 patients with breast cancer with expanded autologous PBPC, with 11 patients receiving expanded PBPC as their sole hematopoietic cell source. In these studies, the CD34+ cells were selected and cultured for 10 days in defined media containing 100 ng/mL each of recombinant human stem cell factor (rhSCF), recombinant human granulocyte colony stimulating factor (rhG-CSF), and recombinant human megakaryocyte growth and developmental factor (rhMGDF) in 1-liter Teflon bags at 20,000 to 50,000 cells/mL. After culture the cells were washed and reinfused after high-dose chemotherapy followed by daily administration of rhG-CSF....","PeriodicalId":51233,"journal":{"name":"Cell Preservation Technology","volume":"4 1","pages":"4-8"},"PeriodicalIF":0.0,"publicationDate":"2006-04-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/CPT.2006.4.4","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"60913333","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}
In life sciences, an increasing number of techniques allow researchers to obtain as much data as possible from biologic specimens. One of the major problems of effective tissue preservation is to retain good tissue morphology and to preserve the nucleotide (DNA/RNA) and protein content of a sample. Effective methods to preserve these products are available; however, their multipurpose application is limited. With the recent introduction of the formaldehyde-releasing agent dimethyldimethylolhydantoin (DMDM-Hydantoin) as an alternative in tissue preservation we were interested in its ability to preserve the RNA and proteins of tissue samples. To test the effect of fluid preservatives on proteins, we used tissue of transgenic embryonic mice that express the β-galactosidase enzyme. Total RNA was isolated with TRIzol® (Invitrogen, Breda, The Netherlands) reagent to analyze the effect of the preservation on RNA. The effectiveness of DMDM-Hydantoin (5%) was tested against more widely used preservatives including...
在生命科学领域，越来越多的技术使研究人员能够从生物标本中获得尽可能多的数据。有效组织保存的主要问题之一是保持良好的组织形态和保存样本的核苷酸(DNA/RNA)和蛋白质含量。有有效的方法来保存这些产品;然而，它们的多用途应用是有限的。随着甲醛释放剂二甲基二甲基酰海丹托因(DMDM-Hydantoin)作为组织保存的替代方法的引入，我们对其保存组织样品的RNA和蛋白质的能力感兴趣。为了测试液体防腐剂对蛋白质的影响，我们使用表达β-半乳糖苷酶的转基因小鼠胚胎组织。用TRIzol®(Invitrogen, Breda, The Netherlands)试剂分离总RNA，分析保存对RNA的影响。DMDM-Hydantoin(5%)对更广泛使用的防腐剂的有效性进行了测试，包括…
{"title":"A Preservation Method Supporting Multipurpose Analysis of Long-stored Samples","authors":"D. Molin, and K V Dam","doi":"10.1089/CPT.2006.4.46","DOIUrl":"https://doi.org/10.1089/CPT.2006.4.46","url":null,"abstract":"In life sciences, an increasing number of techniques allow researchers to obtain as much data as possible from biologic specimens. One of the major problems of effective tissue preservation is to retain good tissue morphology and to preserve the nucleotide (DNA/RNA) and protein content of a sample. Effective methods to preserve these products are available; however, their multipurpose application is limited. With the recent introduction of the formaldehyde-releasing agent dimethyldimethylolhydantoin (DMDM-Hydantoin) as an alternative in tissue preservation we were interested in its ability to preserve the RNA and proteins of tissue samples. To test the effect of fluid preservatives on proteins, we used tissue of transgenic embryonic mice that express the β-galactosidase enzyme. Total RNA was isolated with TRIzol® (Invitrogen, Breda, The Netherlands) reagent to analyze the effect of the preservation on RNA. The effectiveness of DMDM-Hydantoin (5%) was tested against more widely used preservatives including...","PeriodicalId":51233,"journal":{"name":"Cell Preservation Technology","volume":"4 1","pages":"46-50"},"PeriodicalIF":0.0,"publicationDate":"2006-04-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/CPT.2006.4.46","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"60913415","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}
Shijun Zhu, Kamran Jamil, Xiaocui Ma, J. Crowe, A. E. Oliver
CANARY cells are genetically engineered murine B cells that serve as a rapid detection system for various infectious pathogens. We are attempting to produce a stable dehydrated cellular product to make this system practical. A major source of damage to these cells during drying was identified as apoptosis. Trehalose, which protects mammalian cells during drying, was investigated with regard to its effect on apoptosis. B cells were loaded with trehalose and vacuum-dried. Trehalose reduced apoptosis during drying and rehydration, and when used in combination with a pan-caspase inhibitor OPH-109, the degree of apoptotic cell loss was further diminished. In this case, viability following rehydration reached 70%–80%. Surprisingly, trehalose alone blocked apoptotic cell death better than OPH-109 alone (45% versus 70% total apoptotic cells for trehalose- or OPH-109–treated samples, respectively, after drying to 0.3 g H2O/g dry weight). Nevertheless, optimal viability was achieved when the cells were loaded and d...
CANARY细胞是经过基因工程改造的小鼠B细胞，可作为各种感染性病原体的快速检测系统。我们正在尝试生产一种稳定的脱水细胞产品，使该系统实用。这些细胞在干燥过程中受损的主要原因是细胞凋亡。海藻糖具有保护哺乳动物细胞在干燥过程中的作用，研究了海藻糖对细胞凋亡的影响。B细胞装载海藻糖并真空干燥。海藻糖在干燥和复水化过程中减少凋亡，与泛半胱天冬酶抑制剂opho -109联合使用时，凋亡细胞损失程度进一步降低。在这种情况下，补液后的存活率达到70%-80%。令人惊讶的是，单独使用海藻糖比单独使用opho -109更好地阻止凋亡细胞死亡(在干燥至0.3 g H2O/g干重后，海藻糖或opho -109处理的样品中，凋亡细胞总数分别为45%和70%)。然而，当细胞被加载和加载时，达到了最佳的生存能力。
{"title":"Protection of CANARY Cells After Drying and Rehydration Correlates with Decrease in Apoptotic Cell Death","authors":"Shijun Zhu, Kamran Jamil, Xiaocui Ma, J. Crowe, A. E. Oliver","doi":"10.1089/CPT.2006.4.67","DOIUrl":"https://doi.org/10.1089/CPT.2006.4.67","url":null,"abstract":"CANARY cells are genetically engineered murine B cells that serve as a rapid detection system for various infectious pathogens. We are attempting to produce a stable dehydrated cellular product to make this system practical. A major source of damage to these cells during drying was identified as apoptosis. Trehalose, which protects mammalian cells during drying, was investigated with regard to its effect on apoptosis. B cells were loaded with trehalose and vacuum-dried. Trehalose reduced apoptosis during drying and rehydration, and when used in combination with a pan-caspase inhibitor OPH-109, the degree of apoptotic cell loss was further diminished. In this case, viability following rehydration reached 70%–80%. Surprisingly, trehalose alone blocked apoptotic cell death better than OPH-109 alone (45% versus 70% total apoptotic cells for trehalose- or OPH-109–treated samples, respectively, after drying to 0.3 g H2O/g dry weight). Nevertheless, optimal viability was achieved when the cells were loaded and d...","PeriodicalId":51233,"journal":{"name":"Cell Preservation Technology","volume":"4 1","pages":"67-77"},"PeriodicalIF":0.0,"publicationDate":"2006-04-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/CPT.2006.4.67","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"60914173","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}
In recent years, ice-free cryopreservation by vitrification has been demonstrated to provide superior preservation of tissues compared with conventional freezing methods. To date, this has been accomplished almost exclusively for small model systems, whereas cryopreservation of large tissue samples-of a clinically useful size-continues to be hampered by thermomechanical effects that compromise the structure and function of the tissue. Reduction of mechanical stress is an integral condition of successful cryopreservation of large specimens. The current study focuses on the impact of sample size on both the physical events, observed by cryomacroscopy, and on the outcome on tissue function. To this end, the current study sought to address the question of functional recovery of vitrified carotid artery segments, processed as either artery rings (3-4 mm long) or segments (25 mm long) as selected models; the latter model represents a significant increase in sample size for evaluating the effects of vitrification. Tissue vitrification using an 8.4 M cryoprotectant cocktail solution (VS55) was achieved in 1-ml samples by imposing either a high (50-70 °C/min) or a low (2-3 °C/min) cooling rate, between -40°C and -100°C, and a high rewarming rate between -100°C and -40°C. Following cryoprotectant removal, the artery segments were cut into 3 to 4-mm rings for function testing on a contractility apparatus by measuring isometric responses to four agonist and antagonists (norepinephrine, phenylepinephrine, calcium ionophore, and sodium nitroprusside). In addition, nonspecific metabolic function of the vessel rings was determined using the REDOX indicator alamarBlue. Contractile function in response to the agonists norepinephrine and phenylepinephrine was maintained at the same level (350%) for the segments as for the rings, when compared with noncryopreserved control samples. Relaxation in response to the antagonists calcium ionophore and sodium nitroprusside was maintained at between 75% and 100% of control levels, irrespective of cooling rate or sample size. No evidence of macroscopic crystallization or fractures was observed by cryomacroscopy at the above rates in any of the samples. In conclusion, this study verifies that the rate of cooling and warming can be reduced from our baseline vitrification technique such that the function of larger tissue samples is not significantly different from that of smaller blood vessel rings. This represents a step toward the goal of achieving vitreous cryopreservation of large tissue samples without the destructive effect of thermal stresses.
{"title":"Vitrification of Carotid Artery Segments: An Integrated Study of Thermophysical Events and Functional Recovery Toward Scale-Up for Clinical Applications.","authors":"S Baicu, M J Taylor, Z Chen, Y Rabin","doi":"10.1089/cpt.2006.9994","DOIUrl":"10.1089/cpt.2006.9994","url":null,"abstract":"<p><p>In recent years, ice-free cryopreservation by vitrification has been demonstrated to provide superior preservation of tissues compared with conventional freezing methods. To date, this has been accomplished almost exclusively for small model systems, whereas cryopreservation of large tissue samples-of a clinically useful size-continues to be hampered by thermomechanical effects that compromise the structure and function of the tissue. Reduction of mechanical stress is an integral condition of successful cryopreservation of large specimens. The current study focuses on the impact of sample size on both the physical events, observed by cryomacroscopy, and on the outcome on tissue function. To this end, the current study sought to address the question of functional recovery of vitrified carotid artery segments, processed as either artery rings (3-4 mm long) or segments (25 mm long) as selected models; the latter model represents a significant increase in sample size for evaluating the effects of vitrification. Tissue vitrification using an 8.4 M cryoprotectant cocktail solution (VS55) was achieved in 1-ml samples by imposing either a high (50-70 °C/min) or a low (2-3 °C/min) cooling rate, between -40°C and -100°C, and a high rewarming rate between -100°C and -40°C. Following cryoprotectant removal, the artery segments were cut into 3 to 4-mm rings for function testing on a contractility apparatus by measuring isometric responses to four agonist and antagonists (norepinephrine, phenylepinephrine, calcium ionophore, and sodium nitroprusside). In addition, nonspecific metabolic function of the vessel rings was determined using the REDOX indicator alamarBlue. Contractile function in response to the agonists norepinephrine and phenylepinephrine was maintained at the same level (350%) for the segments as for the rings, when compared with noncryopreserved control samples. Relaxation in response to the antagonists calcium ionophore and sodium nitroprusside was maintained at between 75% and 100% of control levels, irrespective of cooling rate or sample size. No evidence of macroscopic crystallization or fractures was observed by cryomacroscopy at the above rates in any of the samples. In conclusion, this study verifies that the rate of cooling and warming can be reduced from our baseline vitrification technique such that the function of larger tissue samples is not significantly different from that of smaller blood vessel rings. This represents a step toward the goal of achieving vitreous cryopreservation of large tissue samples without the destructive effect of thermal stresses.</p>","PeriodicalId":51233,"journal":{"name":"Cell Preservation Technology","volume":"4 4","pages":"236-244"},"PeriodicalIF":0.0,"publicationDate":"2006-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2180387/pdf/nihms35598.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"27215111","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
The potential for use of human mesenchymal stem cells in regenerative medicine has been widely discussed, because these cells are capable of differentiating into bone, muscle, cartilage, adipose, a...
{"title":"Arbutin Enhances Recovery and Osteogenic Differentiation in Dried and Rehydrated Human Mesenchymal Stem Cells","authors":"Kamran Jamil, J. Crowe, F. Tablin, A. E. Oliver","doi":"10.1089/CPT.2005.3.244","DOIUrl":"https://doi.org/10.1089/CPT.2005.3.244","url":null,"abstract":"The potential for use of human mesenchymal stem cells in regenerative medicine has been widely discussed, because these cells are capable of differentiating into bone, muscle, cartilage, adipose, a...","PeriodicalId":51233,"journal":{"name":"Cell Preservation Technology","volume":"3 1","pages":"244-255"},"PeriodicalIF":0.0,"publicationDate":"2005-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/CPT.2005.3.244","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"60912457","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}
Laura Musacchio, N. Greppi, R. Marchesi, P. Rebulla
Plasma for clinical use should be frozen and stored in this state until use. In this study we revised our standard procedure for the cryopreservation of plasma to verify its compliance to Italian, European, and American norms, which require a specific cooling rate. To this aim, we performed three tests. In test 1 we recorded the temperature of the freezing chamber of our mechanical blast freezer; in tests 2 and 3 we recorded and certified with thermocouples the freezing curve of plasma units frozen without and with protective packaging (soft polyethylene bag and cardboard box). In test 2 the freezer was loaded in a single operation, whereas in test 3 we simulated the ordinary loading operation that requires the opening of the loading door on two occasions, which decreases thermal efficiency. The validation of the cooling system, which included a thermal analysis, allowed us to characterize the critical steps of the plasma freezing process. The thermal analysis was conducted under ideal conditions in order...
{"title":"Validation of a Mechanical Freezer Used for the Cryopreservation of Fresh Frozen Plasma: Thermodynamic and Heat Transfer Evaluations","authors":"Laura Musacchio, N. Greppi, R. Marchesi, P. Rebulla","doi":"10.1089/CPT.2005.3.223","DOIUrl":"https://doi.org/10.1089/CPT.2005.3.223","url":null,"abstract":"Plasma for clinical use should be frozen and stored in this state until use. In this study we revised our standard procedure for the cryopreservation of plasma to verify its compliance to Italian, European, and American norms, which require a specific cooling rate. To this aim, we performed three tests. In test 1 we recorded the temperature of the freezing chamber of our mechanical blast freezer; in tests 2 and 3 we recorded and certified with thermocouples the freezing curve of plasma units frozen without and with protective packaging (soft polyethylene bag and cardboard box). In test 2 the freezer was loaded in a single operation, whereas in test 3 we simulated the ordinary loading operation that requires the opening of the loading door on two occasions, which decreases thermal efficiency. The validation of the cooling system, which included a thermal analysis, allowed us to characterize the critical steps of the plasma freezing process. The thermal analysis was conducted under ideal conditions in order...","PeriodicalId":51233,"journal":{"name":"Cell Preservation Technology","volume":"3 1","pages":"223-228"},"PeriodicalIF":0.0,"publicationDate":"2005-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/CPT.2005.3.223","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"60912337","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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