Dendritic cells (DCs) and mast cells (MCs) are key players of the immune system, often coming in close proximity in peripheral tissues. The interplay of these cells is, however, still poorly understood, especially with regards to human cells. The reason for that is the absence of a well established in vitro human cell-based study system that would allow a simultaneous preparation of both cell types. In this study, we show a method for simultaneous generation of DCs and MCs from CD34+ stem cell progenitors that were isolated from the non-adherent fraction of non-mobilized peripheral blood mononuclear cells of healthy donors. We observed that combining stem cells factor (SCF), IL-3 and GM-CSF in serum-free StemPro-34 medium allowed CD34+ cells isolated from an equivalent of 450 ml of peripheral blood to expand to 10-92 × 106 cells after 7 weeks of culturing. These cultures comprised of 6-53% of DCs and 1-21% of MCs as determined by the expression of, respectively, CD11c/HLA-DR or CD117/FcεRI. The DCs were CD1a-CD14-, did not express co-stimulatory molecules CD80 and CD83 and chemokine receptor CCR7. However, the DCs expressed co-stimulatory molecule CD86, and had a capacity to uptake dextran, phagocyte latex particles and induce alloreactivity. MCs, on the other hand, degranulated after crosslinking of FcεRI-bound IgE as determined by the externalization of CD107b. Collectively, our data show that CD34+-derived human DCs and MCs can be generated in a single culture using CD34+ cells isolated from non-mobilized human peripheral blood and that this method may allow ex vivo studies on DC-MC interplay in human system.

• MeSH
• antigeny CD34 metabolismus   MeSH
• buněčná diferenciace MeSH
• degranulace buněk imunologie   MeSH
• dendritické buňky imunologie   metabolismus   MeSH
• dextrany imunologie   MeSH
• faktor růstu kmenových buněk metabolismus   MeSH
• faktor stimulující granulocyto-makrofágové kolonie metabolismus   MeSH
• kmenové buňky z periferní krve fyziologie   MeSH
• kultivační média bez séra metabolismus   MeSH
• leukocyty mononukleární MeSH
• lidé MeSH
• mastocyty imunologie   MeSH
• mezibuněčná komunikace imunologie   MeSH
• primární buněčná kultura metody   MeSH
• průtoková cytometrie metody   MeSH
• rekombinantní proteiny metabolismus   MeSH
• separace buněk metody   MeSH
• vrstva buffy coat cytologie   MeSH
• zdraví dobrovolníci pro lékařské studie MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

The c-myb proto-oncogene and its oncogenic derivative v-mybAMV encode transcriptional regulators engaged in the commitment of hematopoietic cells. While the c-Myb protein is important for the formation and differentiation of various progenitors, the v-MybAMV oncoprotein induces in chicks a progression and transformation of the single (monoblastic) cell lineage. Here we present the first evidence of cell fate-directing abilities of c-Myb and v-MybAMV proteins in avian neural crest (NC), where both proteins determine melanocytogenesis. The increased concentration of c-Myb induces progression into dendritic melanocytes and differentiation. The v-myb oncogene converts essentially all NC cells into melanocytes and causes their transformation. Both Myb proteins activate in NC cells expression of the c-kit gene and stem cell factor c-Kit signaling--one of the essential pathways in melanocyte development. These observations suggest that the c-myb-c-kit pathway represents a common regulatory scheme for both hematopoietic and neural progenitors and establishes a novel experimental model for studies of melanocytogenesis and melanocyte transformation.

• MeSH
• buněčná diferenciace MeSH
• crista neuralis cytologie   metabolismus   MeSH
• DNA primery genetika   MeSH
• faktor růstu kmenových buněk genetika   metabolismus   MeSH
• financování organizované MeSH
• geny myb MeSH
• kuřecí embryo MeSH
• melanocyty cytologie   metabolismus   MeSH
• onkogenní proteiny v-myb genetika   metabolismus   MeSH
• protoonkogenní proteiny c-kit genetika   metabolismus   MeSH
• protoonkogenní proteiny c-myb genetika   metabolismus   MeSH
• sekvence nukleotidů MeSH
• signální transdukce MeSH
• zvířata MeSH
• Check Tag
• kuřecí embryo MeSH
• zvířata MeSH