Supplementary material

Supplementary material

Files in this Data Supplement:

  • Supplementary table and methods - Table: Specific primers for Real-Time RT-PCR amplification Method: Immunofluorescence staining
  • Supplementary figure 1 - Flow cytometry analysis of murine BMMSCs. Time course of cell size and granularity analyzed by flow cytometry. Forward scatter (FS) versus side scatter (SS) plot of whole bone marrow adherent cells (A) and CD11b-CD45-CD31- bone marrow adherent cells (B). Data shown are from one representative of three independent experiments.
  • Supplementary figure 2 - Real-time PCR of stem cell marker and oncogenic marker expression by 2h-BMMSCs, 9d-BMMSCs and long-term cultured BMMSCs. (A): The expression of stem cell markers Oct3/4, Nanog, Sox2, SSEA-1 and Rex-1 were further downregulated during long-term cultivation. (B): The expression of oncogenic marker c-Myc was further upregulated during long-term cultivation. The expression level is normalized to the level of GAPDH.
  • Supplementary figure 3 - Real-time PCR of type I procollagen, α-SMA, TGF-β and c-Myc of 2h-BMMSCs and 9d-BMMSCs. (A, B): 9d-BMMSCs showed markedly higher expression of type I procollagen and α-SMA compared with 2h-BMMSCs. The expression level is normalized to the level of GAPDH. (C): 9d-BMMSCs showed higher expression of TGF-β compared with 2h-BMMSCs. To compare the levels of TGF-β production in the fixed number of donor cells between 2h-BMMSCs and 9d-BMMSCs, the PCR data is again normalized to the level of GAPDH, followed by adjustment to the amount of one cell. (D): 9d-BMMSCs showed higher expression of c-Myc compared with 2h-BMMSCs. The expression level is normalized to the level of GAPDH. 2h-BMMSCs and 9d-BMMSCs were harvested from 10-12 femurs and tibias of 5-6 mice, respectively. Data are expressed as mean�SD. *p <_0.05. xmlns:abbreviations="urn:x-prefix:abbreviations" abbreviations:_="abbreviations:_" alpha-sma="alpha-sma" alpha-smooth="alpha-smooth" muscle="muscle" actin="actin" tgf-beta="tgf-beta" transforming="transforming" growth="growth" factor-beta.--="factor-beta.--" end="end" desc="desc" supplemental_figure_3.jpg="supplemental_figure_3.jpg" _--="_--">
  • Supplementary figure 4 - Reduction of type I collagen synthesis in bleomycin-injected lungs by administration of 2h-BMMSCs and 9d-BMMSCs. (A) Immunofluorescence staining of type I collagen in lungs 10 days after injection of bleomycin followed by 2h-BMMSCs or 9d-BMMSCs transfer (red, type I collagen; blue, nucleus). Magnification 100X. Scale bars, 200µm. Values in (B) give the ratios of the fluorescence intensities of type I collagen to TOPROIII. Although there was no statistically significant difference, the fluorescence intensities of type I collagen was downregulated in mice receiving 2h-BMMSCs or 9d-BMMSCs compared with mice treated with PBS alone. Data are expressed as mean�SD. Comparisons between the two groups were analyzed using paired Student�s t test.
  • Supplementary figure 5 - Evaluation of fibrotic changes by Ashcroft scale in the murine lungs. The Ashcroft scores were measured in the lung H&E section. The severity of fibrotic changes in each section was assessed as the mean score of severity in the observed microscopic fields. Thirty fields per section were analyzed. Data are expressed as mean±SD. **p <_0.01.-- end="end" desc="desc" supplemental_figure_5.jpg="supplemental_figure_5.jpg" _--="_--">