After the obtention of dendritic cells derived from monocytes (GMCSF+IL-4), we are trying to obtain Langerhans cells from the same source. The addition of TGFB does not allow us to obtain very clear results.
Hi Olivier,
Langerhans cells typically self-renew within the skin, so differentiation from blood- or BM-derived monocytes is not common in vivo. However they can be reconstituted from the blood if sufficiently depleted. FYI, there are Langerin+ dermal dendritic cells (they are BM-renewed) and they are NOT Langerhans cells. The two populations are actually dichotomous in some situations. For example, Langerhans cells suppress allergic contact dermatitis, dermal Langerin+ cells promote it. Langerhans cells skew T cell responses toward Th17, Langerin+ dermal dendritic cells toward Th1. In my opinion, if you start with monocytes, you're more likely to get the dermal cells. If this doesn't matter to you, then you can ignore everything I just said. I'm sorry, I don't have the cytokines for in vitro differentiation, but Langerhans cells are highly dependent on TGFb. One more thing - they're pretty easy to isolate out of skin, so maybe you don't need to differentiate them?
The addition of IL-4 prevents macrophage differentiation but can also inhibit Langerhan cell differentiation. Try with GM-CSF+TGFb +TNFa .
Hi Olivier,
Langerhans cells typically self-renew within the skin, so differentiation from blood- or BM-derived monocytes is not common in vivo. However they can be reconstituted from the blood if sufficiently depleted. FYI, there are Langerin+ dermal dendritic cells (they are BM-renewed) and they are NOT Langerhans cells. The two populations are actually dichotomous in some situations. For example, Langerhans cells suppress allergic contact dermatitis, dermal Langerin+ cells promote it. Langerhans cells skew T cell responses toward Th17, Langerin+ dermal dendritic cells toward Th1. In my opinion, if you start with monocytes, you're more likely to get the dermal cells. If this doesn't matter to you, then you can ignore everything I just said. I'm sorry, I don't have the cytokines for in vitro differentiation, but Langerhans cells are highly dependent on TGFb. One more thing - they're pretty easy to isolate out of skin, so maybe you don't need to differentiate them?
I would imagine that co-culturing them with a coated plate of dermal fibroblasts and TSLP would give you langerhan-like cells, I'm unsure if you have BMD monocytes or from the peripheral blood. In theory, I would think PBMC would be better to obtain circulating monocytes ready to infiltrate the skin; however, are those then langerhan cells?
I agree with John that isolating from the skin would be the best, there are perhaps mechanisms to expand them in the literature other than simply IL-4 and GM-CSF.
References:
http://www.jacionline.org/article/S0091-6749(07)00118-2/abstract
To obtain DC from PBMC the best cytokines are GM-CSF + IL-4. Although some researchers believe that IL-13+ GMCSF affect on DC difrentiation even more efficient than IL-4.
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