Experimental attempt to produce mRNA transfected dendritic cells derived from enriched CD34+ blood progenitor cells
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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.
KeywordsCD34+ cell derived dendritic cells mRNA transfected Cancer vaccines Norepinephrine
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- Maria R. Motta, Castellani M., Rizzi S., Curti A., Gubinelli F., Fogli M., et al., Generation of dendritic cells from CD14+ monocytes positively selected by immunomagnetic adsorption for multiple myeloma patients enrolled in a clinical trial of anti-idiotype vaccination., Br. J. Hemetol., 2003, 121, 240–250CrossRefGoogle Scholar
- Ferlazzo G., Wesa A., Wie W., Galy A., Dendritic Cells generated either from D34+ progenitor cells or from monocytes differ in their ability to activate antigen-specific CD8+ T cells, J. Immmunol., 1999, 163, 3597–3604Google Scholar
- Luft T., Pang K., Thomas E., Bradley C., Savoia H., Trapani J., et al., A serum-free culture model for studying the differentiation of human dendritic cells from adult CD34+ progenitor cells, Exp. Hematology, 1998, 26, 489–500Google Scholar
- Kowalkowski K., Alzona M., Aono F., Van Epps D., Vachula M., Ex Vivo Generation of Dendritic Cells from CD34+ Cells in Gas-Permeable containers Under Serum-Free Conditions, J. Hematotherapy, 1998, 7, 403–411Google Scholar
- Kvalheim G., Clinical use of enriched CD34+ cells, Blood Therapies in Medicine, 2002, 2, 57–64Google Scholar