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Biotechnology and Applied Biochemistry (2001) 34, (161–166) (Printed in Great Britain)

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Efficient cryopreservation of dendritic cells transfected with cDNA of a tumour antigen for clinical application

Gabriele Pecher*†¹, Thomas Schirrmann*†, Lothar Kaiser‡ and Jörg A. Schenk†§

*Humboldt-University Berlin, Charité Campus Mitte, AG Molecular Gene- and Immunotherapy, Hessische Str. 3–4, 10115 Berlin, Germany, †Max-Delbrück-Center for Molecular Medicine, Berlin, Germany, ‡Immunological Laboratory Hannover GmbH, Hannover, Germany, and §Department of Biotechnology, University of Potsdam, Golm, Germany

Key words: antigen-presenting cell, gene transfer, mucin.

Abbreviations used: DC, dendritic cell; GMP, good manufacturing practice; PBMC, peripheral blood mononuclear cell; rhuGM-CSF, recombinant human granulocyte/macrophage colony-stimulating factor; rhuIL-4, recombinant human interleukin 4; DMEM, Dulbecco's modified Eagle's medium; HLA, human leucocyte antigen.

¹To whom correspondence should be addressed, at Humboldt-University Berlin (e-mail gabriele.pecher@charite.de).

Dendritic cells (DCs) are the most potent antigen-presenting cells of the immune system and are currently being investigated in clinical applications as cancer vaccines. An efficient cryopreservation method would greatly contribute to their use in clinical trials. We have established a method for freezing of DCs derived from peripheral blood mononuclear cells using the plasma expander Gelifundol^®. This enabled us to reduce the concentration of the toxic DMSO to 5%. The method could be performed without the addition of fetal calf serum or any other serum. After freezing, the viability of the DCs was 90%. The cells exhibited all the phenotypic characteristics (CD11c+, HLA-DR+, CD80+, CD83+, CD86+) of DCs, as tested by flow cytometry. Cells transfected with cDNA for the tumour antigen mucin expressed this protein on their surfaces in the same manner as before freezing. The stimulating capacity of a mixed lymphocyte culture was also preserved. These findings offer an efficient method for the cryopreservation of DCs for use in clinical trials.

Received 20 May 2001/15 August 2001; accepted 17 August 2001

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