Shinde, P. and Khan, N. and Melinkeri, S. and Kale, V. and Limaye, L. (2019) Freezing of dendritic cellswith trehalose as an additive in the conventional freezing medium results inimproved recovery after cryopreservation. Transfusion., 59 (2). pp. 686-696.

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Abstract

BACKGROUND:Dendritic cell (DC) vaccinationinvolves administration of multiple doses.Cryopreservation of tumor antigen–pulsed DCs canprovide a ready to use vaccine source and eliminate theneed of frequent withdrawal of the patient’s blood forvaccine preparation. The aim of this study was to assessthe effect of addition of trehalose in the freezing mediumon the recovery of DCs after cryopreservation.STUDY DESIGN AND METHODS:DCs weregenerated from mononuclear cells from apheresissamples of healthy donors. For long-term storage of6 months, cells were frozen with a rate-controlledprogrammable freezer and stored in liquid nitrogen. Forshort-term storage of 1 month, cells were frozen andstored at−80�C. DCs frozen with Iscove’s ModifiedDulbecco’s Medium + 10% dimethyl sulfoxide + 20%fetal bovine serum served as the control group, while thetest group was additionally supplemented with 50μg/mLof trehalose. After revival of control and test DCs, theywere assessed for viability, morphology, phenotype, andfunctions.RESULTS:The addition of trehalose to the conventionalfreezing medium helped to preserve the viability andfunctionality of DCs better than dimethyl sulfoxide alonein both long- and short-term cryopreservation. Trehalosealso protected the mitochondrial membrane potential andcytoskeleton integrity of DCs, which are necessary fortheir functionality. Mediators of the intrinsic apoptoticpathway like Caspase-9 and Bim-1 were found to be lowin the test.CONCLUSION:Supplementation of conventionalfreezing medium with trehalose results in better quality ofDCs revived after cryopreservation. Thisfinding couldhelp improve DC vaccine preparation for cancerimmunotherapy.

Item Type:	Article
Depositing User:	Mr. Rameshwar Nema
Date Deposited:	14 Feb 2020 04:14
Last Modified:	14 Feb 2020 04:14
URI:	http://nccs.sciencecentral.in/id/eprint/645

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