Determination of residual dimethylsulfoxide in cryopreserved cardiovascular allografts
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Determination of residual dimethylsulfoxide in cryopreserved cardiovascular allografts R. Dı´az Rodrı´guez . B. Van Hoeck . S. De Gelas . F. Blancke . R. Ngakam . K. Bogaerts . R. Jashari
Received: 16 November 2016 / Accepted: 15 December 2016 Ó Springer Science+Business Media Dordrecht 2017
Abstract Dimethylsulfoxide (DMSO) is a solvent which protects the structure of allografts during the cryopreservation and thawing process. However, several toxic effects of DMSO in patients after transplantation of cryopreserved allografts have been described. The aim of this study is to determine the residual DMSO in the cardiovascular allografts after thawing and preparation of cryopreserved allografts for clinical application following guidelines of the European Pharmacopoeia for DMSO detection. Four types of EHB allografts (aortic valve-AV, pulmonary valvePV, descending thoracic aorta-DA, and femoral artery-FA) are cryopreserved using as cryoprotecting solution a 10% of DMSO in medium 199. Sampling is carried out after thawing, after DMSO dilution and after delay of 30 min from final dilution (estimated delay until allograft implantation). After progressive
R. Dı´az Rodrı´guez (&) B. Van Hoeck R. Ngakam R. Jashari European Homograft Bank (EHB), Saint Jean Clinic, Brussels, Belgium e-mail: [email protected] S. De Gelas F. Blancke Laboratoire de Coˆntrole et d’Analyses (LCA), Brussels, Belgium
thawing in sterile water bath at 37–42 °C (duration of about 20 min), DMSO dilution is carried out by adding consecutively 33, 66 and 200 mL of saline. Finally, tissues are transferred into 200 mL of a new physiologic solution. Allograft samples are analysed for determination of the residual DSMO concentration using a validated Gas Chromatography analysis. Femoral arteries showed the most important DMSO reduction after the estimated delay: 92.97% of decrease in the cryoprotectant final amount while a final reduction of 72.30, 72.04 and 76.29% in DMSO content for AV, PV and DA, was found, respectively. The residual DMSO in the allografts at the moment of implantation represents a final dose of 1.95, 1.06, 1.74 and 0.26 mg kg-1 in AV, PV, DA and FA, respectively, for men, and 2.43, 1.33, 2.17 and 0.33 mg kg-1 for same tissues for women (average weight of 75 kg in men, and 60 kg in women). These results are seriously below the maximum recommended dose of 1 g DMSO kg-1 (Regan et al. in Transfusion 50:2670–2675, 2010) of weight of the patient guaranteeing the safety and quality of allografts. Keywords Dimethylsulfoxide Cardiovascular allografts Cryopreservation
K. Bogaerts I-BioStat, KU Leuven, Leuven, Belgium
Introduction
K. Bogaerts Universiteit Hasselt, Hasselt, Belgium
Cryopreservation of cardiovascular tissues is a crucial step in preservation and storage of cardio-vascular
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Cell Tissue Bank
allografts. The structural integrity of the tissue has to be perfectly preserved in order to guarantee the tissue safety, quality and durability. For this purpose, several cryoprotectant solutions are used in cell and tiss
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