Experimental attempt to produce mRNA transfected dendritic cells derived from enriched CD34+ blood progenitor cells

• Authors: Paula Lazarova , Gunnar Kvalheim , Liana Gercheva , Krassimir Metodiev
• Languages of publication: EN

Abstracts

EN

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.

Keywords

EN

CD34+ cell derived dendritic cells; mRNA transfected; Cancer vaccines Norepinephrine

• Publisher: De Gruyter Open
• Journal: Open Medicine
• Year: 2008
• Volume: 3
• Issue: 1
• Pages: 21-28

Dates

published

1 - 3 - 2008

online

1 - 3 - 2008

Contributors

author

Paula Lazarova

author

Gunnar Kvalheim

• Department of Cellular Therapy, University Rikshospital-Radiumhospital, Oslo, Norway

author

Liana Gercheva

• Department of Haematology, Univ. Hospital “St.Marina”, Medical University, Varna, Bulgaria

author

Krassimir Metodiev

• Department of Immunology, Medical University, Varna, Bulgaria

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Identifiers

DOI

10.2478/s11536-007-0072-9