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. 2018 Jul 25;9:1725. doi: 10.3389/fimmu.2018.01725

Figure 3.

Figure 3

Gingival fibroblasts (GFs) are involved in the retention and survival of T and B cells after coculture with peripheral blood mononuclear cells (PBMCs). Overlay of immunohistochemical stainings with 4′,6-diamidino-2-fenylindool (DAPI) and CD3+ or CD20+ staining of (A,B) monocultures of PBMCs and (C,D) GF cocultures with PBMCs after 21 days of culture. (A,C) CD3+ and (B,D) CD20+ stainings (green) were performed to identify T and B cells, respectively. Nuclei are stained with DAPI (blue). GF nuclei are depicted with white arrows. CD3+ cells (A,C) and CD20+ cells (B,D) are depicted with yellow arrows, which are abundantly present in cocultures with GFs. Micrographs are representatives for two independent experiments with five different GF sources. Scale bars represent 100 µM. Proportions of CD3+ (E) and CD20+ (F) cells of total mononuclear cells after 7, 14, and 21 days were quantified from two standardized pictures per well. Significantly more CD3+ and CD20+ cells were identified in GF cocultures. The number of mononuclear cells interacting with TRACP+ cells (G) in PBMC monoculture (white bar) and GF coculture (gray bar). No significant difference (p = 0.0561) between mono- and cocultures was found. n = 5 GF donors, n = 2 buffy coats **p < 0.01, ***<0.001, ****<0.0001.