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Blood, Vol. 110, Issue 9, 3438-3446, November 1, 2007
Multipotent cells can be generated in vitro from several adult human organs (heart, liver, and bone marrow)
Blood Beltrami et al.
110: 3438
Supplemental materials for: Beltrami et al
Files in this Data Supplement:
- Document 1. Supplemental materials and methods (PDF, 107 KB)
- Table S1. RT-PCR primers and product lengths (PDF, 242 KB)
- Table S2. FACS analysis of MASCs (PDF, 242 KB) -
Cell fraction of BM derived, heart derived and liver derived cells, growing in expansion medium, expressing the indicated cell surface marker. Results are indicated as meanħstandard deviation. A Student’s t test was applied in order to assess statistically significant differences in the expression of the tested markers among cell lines derived from the three different tissues. p was considered significant when <0.05.
- Table S3. Multilineage differentiation of single cell derived clones (PDF, 240 KB) -
When differentiation experiments were performed in cells at the 15th generation after single cell seeding, more than 70% of the clones differentiated along the pre-established cell fate (upper part of the table). When multilineage differentiation was performed in single cell derived clones expanded for at least 19 generations, more than 60% of the clones resulted to be multipotent (lower part of the table).
- Table S4. Some of the genes more highly upregulated (A) and downregulated (B) in MASCs with respect to the profiled somatic cell lines and adult tissues (PDF, 461 KB)
- Figure S1. Relation between initial plating density and hMASC expansion (JPG, 181 KB)
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Cardiac derived, liver derived, and bone marrow derived cells were plated on 6 well plates at 1,000, 2,000, 4,000, and 8,000 cells/cm2. The cells were harvested and counted up to ten days after plating. Cell density (expressed as cells per cm2) (A) and fold increase (B) are shown.
- Figure S2. ABCG2 and MDR1 expression (JPG, 56 KB)
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(A) Representative Flow Cytometry dot plots of antigen expression on cells grown in expansion medium. Left plot represents isotype control IgG-staining profile; central plot represents ABCG-2 staining, whereas right plot displays MDR-1 staining. (B) Representative MDR-1 and ABCG2 RTPCR expression analysis on human Heart derived- (hH), Liver derived- (hL) and BM derived- (hBM) cells.
- Figure S3. Tissue specific transcription factor expression (JPG, 191 KB)
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(A) Representative flow-cytometry dot plots of the tested tissue-specific transcription factor expression. (B) Histogram representation of the fraction of the cell population expressing the tested proteins. Data are presented as mean and standard deviation. (C) Representative RT-PCR transcription factor expression analysis on Heart derived- (hH1, hH2 and hH3), Liver derived- (hL1, hL2 and hL3) and Bone Marrow derived- (hBM1, hBM2 and hBM3) cells.
- Figure S4. Multilineage differentiation potential of polyclonal hMASCs (JPG, 403 KB)
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Cardiac derived (left column), liver derived (central column) and BM derived (right column) cells were exposed for 5 to 28 days to differentiation inducing conditions and analyzed thereafter. Cells exposed to an osteogenic medium were positive for the alkaline phosphatase reaction (A, brown staining), and, they were able to produce osteocalcin (B, white fluorescence). Cells exposed for five to fourteen days to a myogenic medium added with bFGF, VEGF and IGF1 expressed increasing amounts of  -sarcomeric actin (C, red fluorescence) and -smooth muscle actin (D, green fluorescence). Cells grown for five to fourteen days in a serum-free medium containing VEGF were decorated with a von Willebrand Factor antibody (E, yellow fluorescence). Cells exposed to FGF-4 and HGF became positive for cytokeratins 18 and 19 (F, purple fluorescence). Cells induced to differentiate towards a neural lineage became positive to GFAP staining (G, brown staining) and NSE antibody (H, cyan fluorescence). Nuclei are depicted by the blue fluorescence of DAPI staining. Scale bars=10µm.
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