Medizinische Universität Graz Austria/Österreich - Forschungsportal - Medical University of Graz
Gewählte Publikation:
Bello-Fernandez, C; Matyash, M; Strobl, H; Pickl, WF; Majdic, O; Lyman, SD; Knapp, W.
Efficient retrovirus-mediated gene transfer of dendritic cells generated from CD34+ cord blood cells under serum-free conditions.
Hum Gene Ther. 1997; 8(14):1651-1658
Doi: 10.1089/hum.1997.8.14-1651
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- Co-Autor*innen der Med Uni Graz
- Strobl Herbert
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- Abstract:
- A retroviral-vector encoding the low affinity nerve growth factor receptor (LNGFR) was used to transduce dendritic cells (DCs) generated from CD34+ cord blood (CB) progenitor cells under serum-free conditions. Transduction efficiency was monitored by flow cytometry (FACS) using a specific monoclonal antibody. Prior to retroviral infections, CD34+ CB cells were stimulated for 60 h in a serum-free medium containing a DC differentiation inducing cytokine cocktail: stem cell factor (SCF), granulocyte/macrophage-colony stimulating factor (GM-CSF), tumor necrosis factor alpha (TNFalpha), and transforming growth factor beta 1 (TGF-beta1). Addition of flt3-ligand (FL) to the aforementioned growth factors significantly enhanced cell expansion (41.7+/-11.5 fold vs. 22.5+/-4.7 fold without FL) and generation of CD1a+ DCs (mean 45.7+/-9.8% vs. 28+/-6.5% without FL, n = 4,p = 0.01). Furthermore, FL significantly increased the proportion of CD1a+LNGFR+ cells (mean 10%+/-4.4% vs. 6%+/-2.4 without FL n = 4, p = 0.03). When serum-free viral supernatants were used to infect DCs progenitors under entirely serum-free conditions and with the most potent cytokine combination, approximately one-third of the CD1a+ DCs generated co-expressed the LNGFR gene. Moreover, the transduced gene was also identified in more mature CD1a+CD80+ and CD1a+CD86+ DCs after 12-14 days of culture. In addition, transduced CD1a+ DCs maintained their functional properties, stimulating allogeneic T cells with similar efficiency as nontransduced CD1a+ DCs. Thus, the serum-free system described allows efficient generation and transduction of CD1a+ DCs derived from CD34+ progenitor cells and may be very useful for future therapeutic applications of DCs.
- Find related publications in this database (using NLM MeSH Indexing)
- Antigens, CD1 - analysis
- Antigens, CD34 - analysis
- Cells, Cultured -
- Culture Media, Serum-Free -
- Cytokines - pharmacology
- Dendritic Cells - chemistry
- Fetal Blood - cytology
- Gene Expression -
- Gene Transfer Techniques -
- Hematopoietic Stem Cells -
- Humans -
- Lymphocyte Activation -
- Membrane Proteins - pharmacology
- Receptors, Nerve Growth Factor - genetics
- Retroviridae - genetics
- T-Lymphocytes - immunology