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. Author manuscript; available in PMC: 2011 Dec 12.
Published in final edited form as: Sci Transl Med. 2010 Nov 10;2(57):57ra82. doi: 10.1126/scitranslmed.3001510

Figure 2. Induction by CpG of CD14+ Human Monocytes to Differentiate into Fibrocyte-like Cells. A.

Figure 2

Experimental scheme for the in vitro differentiation of CD14+ monocytes. B. Photomicrographs of monocytes cultured in serum-free media or serum-free media containing 10 ng/ml TGFβ and stimulated with nothing (a,b), 50 μg/mL non CpG (c,d), 50 μg/mL CpG (e,f), or 50 μg/mL poly IC (g, h) on Day 3. C. qRT-PCR analysis of fibrocyte markers. αSMA gene expression in monocytes cultured for 3 d in serum-free media +/- CpG for 24 h (a). Collagen 1 gene expression in monocytes cultured for 3 d in serum-free media or TGFβ, +/- CpG or poly I:C (b). D. Fluorescent ICC for collagen 1 in monocytes (40× magnification) cultured in serum-free media (a) or TGFβ (b); serum-free media + CpG (c), or TGFβ + CpG (d). Isotype control for monocytes cultured in TGFβ + CpG (e). Representative (n=3) FC for collagen 1 protein as percent of CD14+ cells in CD45+ gate from monocytes cultured in serum-free media or TGFβ, and serum-free media + CpG or TGFβ + CpG (f). E. Forward and side scatter FC of monocytes cultured in serum-free media containing TGFβ (a) or TGFβ + CpG (b). Representative (n=3) FC for CD14 as percent of total cells from monocytes cultured in serum-free media (c) or monocytes cultured in serum-free media containing TGFβ (d) stained with anti-CD14. F. Representative (n=3) FC for CD45 as percent of CD14- cells from monocytes cultured in serum-free media (e) or monocytes cultured in serum-free media containing TGFβ (f) stained with anti-CD45 and gated with respect to CD14 expression. Representative data (n=3) is graphed as percent of CD14+ cells from monocytes cultured in serum-free media (g) and monocytes cultured in serum-free media containing TGFβ (h) stained with anti-CD45 and gated with respect to CD14 expression.