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. 2016 Jan 26;5:e09394. doi: 10.7554/eLife.09394

Figure 2. Mismatched donor derived-fibroblasts engrafted in target tissues of fibrosis.

(A, B) Transplantation scheme of EGFP+-labeled B10.D2 PαS-BMSCs (A) or SP-HSCs (B). Each was co-transplanted into wild-type BALB/c mice along with unlabeled SP-HSCs (A) or PαS-BMSCs (B), respectively. Arrows indicate colocalized cells in yellow (GFP-labeled BMSCs expressed HSP47) in the lacrimal gland and intestine. (C) Syngeneic wild-type B10.D2 SP-HSCs and EGFP+-labeled B10.D2 PαS-BMSC co-transplantation into wild B10.D2 mice. HSP47+ (red) fibroblasts observed in mismatched BALB/c recipients were BMSC-derived, and not HSC cells. Nuclei were counter stained with DAPI (blue). The data in Figure 2AC from two replicate experiments (n = 3 per group). Scale bar, 20 μm. (AC) D, duct. (D) Cultured HSP47+ lacrimal gland fibroblasts after mismatched EGFP+ HSC transplantation were EGFP- (left), while the majority of fibroblasts after mismatched EGFP+ BMSC transplantation were EGFP+ fibroblasts (right, positive cells in yellow) indicating the donor BMSC origin of fibroblasts. (E) Donor–derived EGFP+ cells were observed in the spleen 3 weeks after syngeneic EGFP+ WBMT (left), while engraftment of donor cells were sparse following mismatched WBMT (right), indicating a number of residual host cells remained after mismatched WBMT. Figure 2D,E from representative data of two replicate experiments (n = 2 or 3 per group). Scale bar, D = 50 μm, E = 20 μm. BMSC, bone marrow stromal/stem cells; HSP, heat-shock protein; HSCs, hematopoietic stem cells

DOI: http://dx.doi.org/10.7554/eLife.09394.013

Figure 2—source data 1. Percentage of donor–derived EGFP+ cells in the spleen 3 weeks after EGFP+ WBMT.
Source data for graph in right panel of Figure 2E.
DOI: 10.7554/eLife.09394.014

Figure 2.

Figure 2—figure supplement 1. Mismatched PαS-BMSCs do not induce sclerodermatous fibrosis in recipient nude mice.

Figure 2—figure supplement 1.

(A) Transplantation scheme of B10D.2 (Mismatched) PαS-BMSCs and BALB/c-nu/nu (Syngeneic) SP-HSCs into nu/nu or wild type BALB/c. (B) Mallory staining of the lacrimal gland, salivary gland, and skin in BALB/c-nu/nu (top) and wild-type (bottom) recipient mice lacrimal gland and salivary gland at 3 weeks and skin at 8 weeks after transplantation. Data collected from two replicate experiments (n = 3 per group). Scale bar, 50 μm. *, areas of lymphocytic infiltration and excessive fibrosis in deep blue, double arrow; thickness of epidermis. BMSC, bone marrow stromal/stem cells; HSC, hematopoietic stem cells