Hello,
I generated human immature monocyte-derived DCs from CD14-monocytes using GM-CSF and IL-4 for 6 days.
At D6, I see that all cells highly express CD209 (as compared to control monocytes). However, 40% of the cells also express CD14 (same high level as in monocytes).
What are these CD14+ CD209+ cells ? Are they also DCs ?
Thank you.
With my best regards.
Benjamin Tschumi please see previous answers (https://www.researchgate.net/post/Some_CD14_expression_on_monocyte-derived_DCs2) some cd14 expression is expected and on 7th day their % should decrease.
Rabeah Al-Temaimi Thank you very much. I saw this answer inded. In my case, these cells express CD209 and highly CD14. I was wondering whether these are monocytes, macrophages or cells stuck in between DC and monocytes ?
Yes, CD14+ CD209+ are DC cells
https://www.scienceopen.com/document_file/ff64531d-7da7-49ae-8e88-010096d76e7a/PubMedCentral/ff64531d-7da7-49ae-8e88-010096d76e7a.pdf
Previously, we also received immature monDCs that retained a lot of CD14. Now we use cytokines from R&D (USA) or from SciStore (Russia). Now immature DCs completely lose CD14. The stimulation scheme is usual. Cytokines were added twice (at the beginning of cultivation and on the 3rd day of cultivation) IL-4 at a concentration of 20 ng/ml, GM-CSF at a concentration of 100 ng/ml. Immature DCs are usually harvested on the 7th day. An example of a phenotype is in the file.
Thank you Vladimir Yurevich Talayev . Those CD14+ MoDCs were still usable (for instance in maturation experiments) ?
The “immature” monDCs obtained by us are indeed more mature than freshly isolated blood borne classical DCs (an example of the phenotype of blood DCs enriched by magnetic separation in Fig. 1). "Our" monDСs carry many CD80, while blood borne DCs are practically devoid of CD80. The level of CD86 expression is also higher on monDСs. But working with blood borne DCs is difficult. Isolation is expensive, a lot of blood is needed, and purified DCs spontaneously mature in vitro. Therefore, we often use monDCs to evaluate the effect of MPPs, vaccines, adjuvants, or cytokines on the activation of DCs (transformation of "immature" DCs into "mature" DCs). The picture I posted earlier shows the phenotype of immature monDCs and mature monDCs (after stimulation with IL-1b + IL-6 + TNF-a + prostaglandin E2, 48 h.). You can see the stimulation-induced increase in the expression of all markers except CD14. I am also sending you an example of the effect of well-known vaccines on the expression of CD83 and CD86 on monDCs (Fig. 2).