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Improved cryopreservation of human hepatocytes using a new xeno free cryoprotectant solution 被引量:2
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作者 Mohammed Saliem Frida Holm +5 位作者 Rosita Bergstrm Tengzelius Carl Jorns Lisa-Mari Nilsson Bo-Gran Ericzon Ewa Ellis Outi Hovatta 《World Journal of Hepatology》 CAS 2012年第5期176-183,共8页
AIM:To optimize a xeno-free cryopreservation protocol for primary human hepatocytes.METHODS:The demand for cryopreserved hepatocytes is increasing for both clinical and research purposes.Despite several hepatocyte cry... AIM:To optimize a xeno-free cryopreservation protocol for primary human hepatocytes.METHODS:The demand for cryopreserved hepatocytes is increasing for both clinical and research purposes.Despite several hepatocyte cryopreservation protocols being available,improvements are urgently needed.We first compared controlled rate freezing to polystyrene box freezing and did not find any significant change between the groups.Using the polystyrene box freezing,we compared two xeno-free freezing solutions for freezing of primary human hepatocytes:a new medium(STEM-CELLBANKER,CB),which contains dimethylsulphoxide(DMSO) and anhydrous dextrose,both permeating and non-permeating cryoprotectants,and the frequently used DMSO-University of Wisconsin(DMSOUW) medium.The viability of the hepatocytes was assessed by the trypan blue exclusion method as well as a calcein-esterase based live-dead assay before and after cryopreservation.The function of the hepatocytes was evaluated before and after cryopreservation by assessing enzymatic activity of 6 major cytochrome P450 isoforms(CYPs):CYP1A2,CYP2C9,CYP2C19,CYP2D6,CYP3A4 and CYP3A7.RESULTS:The new cryoprotectant combination preserved hepatocyte viability significantly better than the standard DMSO-UW protocol(P < 0.01).There was no significant difference in viability estimation between both the trypan blue(TB) and the Live-Dead Assay methods.There was a correlation between viability of fresh hepatocytes and the difference in cell viability between CB and DMSO protocols(r 2 = 0.69) using the TB method.However,due to high within-group variability in the activities of the major CYPs,any statistical between-group differences were precluded.Cryopreservation of human hepatocytes using the cryoprotectant combination was a simple and xeno-free procedure yielding better hepatocyte viability.Thus,it may be a better alternative to the standard DMSO-UW protocol.Estimating CYP activities did not seem to be a relevant way to compare hepatocyte function between different groups due to high normal variability between different liver samples.CONCLUSION:The cryoprotectant combination may be a better alternative to the standard DMSO-UW protocol in primary human hepatocyte cryopreservation. 展开更多
关键词 Human HEPATOCYTES VIABILITY CYTOCHROME P540 dimethylsulphoxide CRYOPROTECTANT CRYOPRESERVATION
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Studies on the Nucleophilicity and Scavenge of Superoxide Ion by Cyclic Voltammetry
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作者 Wei Ying\|liang, Dang Xue\|ping, Hu Sheng\|shui College of Chemistry and Molecular Sciences, Wuhan University , Wuhan, 430072, China 《Wuhan University Journal of Natural Sciences》 CAS 2003年第03A期866-870,共5页
Superoxide ion was generated by the electrochemical reduction of oxygen at a platinum electrode in dimethylsulphoxide (DMSO). This work was focused on the nucleophilicity and scavenge of electrogenerated\|superoxide ... Superoxide ion was generated by the electrochemical reduction of oxygen at a platinum electrode in dimethylsulphoxide (DMSO). This work was focused on the nucleophilicity and scavenge of electrogenerated\|superoxide ion by cyclic voltammetry. The nucleophilic displacement reactions of superoxide ion with ethyl acetate and diethyl adipate were discussed and the reason for remarkable influence of diethyl adipate was elucidated. The scavenging activity of ascorbic acid was evaluated and the result allowed the conclusion that the scavenging ability of ascorbic acid is much lower in DMSO than in aqueous phase. UV\|spectrum of electrogenerated superoxide ion in DMSO exhibited a single absorption band with λ max at 275 nm, which certified further that the method of electrogeneration was reliable and superoxide ion was stable in DMSO. 展开更多
关键词 superoxide ion dimethylsulphoxide cyclic voltammetry ascorbic acid
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