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Additional Supporting Information may be found in the online version of this article.

FilenameFormatSizeDescription
sc-12-0412_sm_SupplMethods.pdf100KSupplementary Data
sc-12-0412_sm_SupplResults.pdf76KSupplementary Data
sc-12-0412_sm_SupplFigure1.tif334KSupplemental Figure 1. Bioluminescence and fluorescence imaging (BLI/FRI) for Tg(Fluc-egfp) mice. Colored scale bars represent optical radiance intensity in photons/second/cm2/steridian (P·s−1·cm−2·sr−1).
sc-12-0412_sm_SupplFigure2.tif401KSupplemental Figure 2. Multipotent differentiation of mADSCsFluc+GFP+ in vitro. Osteogenesis was identified using alkaline phosphatase (A) and alizarin red-S staining (B); adipogenesis and chondrogenesis were determined using Oil red-O staining (C) and collagen-II immunohistochemistry staining (D) respectively. Scale bar represents 50 μm (A,B) and 5 μm (C,D).
sc-12-0412_sm_SupplFigure3.tif888KSupplemental Figure 3. In vivo bioluminescence imaging and ex vivo luciferase assay of the engrafted mADSCsFluc+GFP+ in murine ischemic gastrocnemius muscle. (A) Bioluminescence imaging (BLI) tracked the survival and kinetics of engrafted mADSCFluc+GFP+ (1.0×107) in murine ischemic gastrocnemius muscle (n=10 for each group). Quantitative analysis of (B) in vivo Fluc optical signals and (C) ex vivo Fluc enzymatic activity on POD14 (n=5) demonstrated the progressive death of mADSCsFluc+GFP+ following transplantation. Low-dose rapamycin (RapaLD, 50 nM·kg− 1·day−1) treatment also significantly facilitated the mADSCs' survival in the ischemic gastrocnemius muscle. Colored scale bar represents BLI average radiance intensity in P·s− 1·cm−2·sr−1. Error bars represent mean ± SD. *P<0.01 vs. PBS, #P<0.001 vs. ADSC.
sc-12-0412_sm_SupplFigure4.tif538KSupplemental Figure 4. Immunofluorescence assay of angiogenesis. Confocal images of adductor muscle frozen sections double stained with endothelial marker CD31 (PECAM−1, red) and 4′,6-diamidino−2-phenylindole (DAPI, blue) on POD14. Scale bar represents 50 μm.
sc-12-0412_sm_SupplFigure5.tif191KSupplemental Figure 5. Expression of inflammatory cytokines in the adductor and gastrocnemius muscles differ after the induction of ischemia. Levels of (A) interleukin−1β (IL−1β), (B) tumor necrosis factor-alpha (TNF-α), (C) IL-6 and (D) IL−10 in the left adductor and gastrocnemius muscle were measured using ELISA on post-operative day1 (POD1), POD7 and POD14. Results are presented for the left adductor muscle in black columns, and the left gastrocnemius muscle in grey columns. Following the induction of ischemia, IL−1β/TNF-α levels are significantly higher, while IL−10 level is significantly lower in the gastrocnemius muscle than the adductor muscle on POD7 and POD14. Error bars represent mean ± SD. *P<0.01 vs. Adductor.
sc-12-0412_sm_SupplFigure6.tif242KSupplemental Figure 6. Matrigel plug assay of infiltrating cells. Fluorescence activated cell sorting (FACS) and flow cytometry analysis of cells infiltrating into the cellular graft site (Matrigel plug) in ischemic hindlimb, with quadrant percentage. The Ly-6Chigh cells (black square, left panels) were sorted to be further analyzed using FACscan cytometer for CD115/F4-80 (right panels). Macrophages were defined as Ly-6Chigh/CD115+/F4-80+ cells; neutrophils were defined as Ly-6Ghigh/Ly- 6Cdull/CD115neg cells.
sc-12-0412_sm_SupplFigure7.tif314KSupplemental Figure 7. Apoptosis assay of mADSCs over hypoxia/reoxygenation in vitro. Fluorescence microscopy showed the TUNEL-positive apoptotic cells (green) following hypoxia/reoxygenation. n=20 random fields. Scale bar represents 10 μm.
sc-12-0412_sm_SupplTable1.pdf30KSupplementary Table 1.
sc-12-0412_sm_SupplTable2.pdf71KSupplementary Table 2.

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