Experimental attempt to produce mRNA transfected dendritic cells derived from enriched CD34+ blood progenitor cells |
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Authors: | Paula Lazarova Gunnar Kvalheim Liana Gercheva Krassimir Metodiev |
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Institution: | (1) Department of Cellular Therapy, University Rikshospital-Radiumhospital, Oslo, Norway;(2) Clinical laboratory, District Hospital “St. Anna”, Varna, Bulgaria;(3) Department of Haematology, Univ. Hospital “St.Marina”, Medical University, Varna, Bulgaria;(4) Department of Immunology, Medical University, Varna, Bulgaria |
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Abstract: | It Peripheral blood progenitor enriched CD34+ cells (PBPC) are rather often used as stem cell background in cancer patients
following high dose therapy. Keeping in mind that precursor dendritic cells (DCs) originate from haematopoietic progenitor
cells, purified CD34+ cells might also serve as starting cells for ex-vivo production of DC. The aim of the present study
is to develop a clinical grade procedure for ex-vivo production of DC derived from enriched CD34+ cells. Various concentrations
of CD34+ cells were grown in gas-permeable Teflon bags with different serum-free and serum-containing media supplemented with
GM-CSF, IL-4, TNF-a, SCF, Flt-3L and INF-a. Serum-free CellGroSCGM medium for 7 days followed by CellGroDC medium in 7 days
gave equal results as serum-containing medium. Following incubation, the cultured cells containing immature DCs were concentrated
and transfected with tumour mRNA from human prostate cancer cell lines employing a highly efficient electroporation procedure.
Thawed transfected DCs were able to elicit primary T-cell responses in vitro against antigens encoded by the prostate cancer
mRNA as shown by ELISPOT assay using mock-transfected DCs as control. The results of our study show that frozen enriched CD34+
cells can be an alternative and efficient source for production of DCs for therapeutic purpose. |
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Keywords: | CD34+ cell derived dendritic cells mRNA transfected Cancer vaccines Norepinephrine |
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