( C) Tri-lineage differentiation capacity of hBMSC-Luc/GFP was demonstrated by the positive formation of a cartilage-like matrix (Alcian Blue, scale bar 100 μm), ( D) mineral deposits indicative of osteogenic tissue (Alizarin Red stain, scale bar 100 μm), and ( E) oil droplet formation indicative of adipogenesis (Oil Red O stain, scale bar 50 μm). ( A) hBMSC-Luc/GFP retained spindle-like morphology, and ( B), were GFP+ (scale bar 100 μm). Characterization of donor 2 transduced hBMSC-Luc/GFP at passage 4. Statistical analysis using a Pearson co-efficient demonstrated a linear relationship between cell number and bioluminescence. Error bars represent the standard deviation of each point. ( G and H) Titration of hBMSC-Luc/GFP demonstrated that luciferase activity and resultant bioluminescence was linearly proportional to cell number. Isotype staining is in shaded gray peaks. (F) Flow cytometry analysis demonstrated that hBMSC-Luc/GFP (green peak) were positive for GFP, CD90, CD105, CD44, CD146, CD73 and negative for CD45. (C) Tri-lineage differentiation capacity of hBMSC-Luc/GFP was demonstrated by the positive formation of a cartilage-like matrix (Alcian Blue, scale bar 100 μm), (D) mineral deposits indicative of osteogenic tissue (Alizarin Red stain, scale bar 100 μm), and (E) oil droplet formation indicative of adipogenesis (Oil Red O stain, scale bar 50 μm). (A) hBMSC-Luc/GFP retained spindle-like morphology, and (B) were GFP + (scale bar 100 μm). Characterization of donor 1 transduced hBMSC-Luc/GFP at passage 4. hBMSC were positive for CD73, CD90, CD105, CD44, CD146, and negative for CD31, CD34, CD45, and HLA-DR. In flow cytometry histograms, isotypes control staining is represented by shaded gray peaks, and stained hBMSC are represented with a black line. (C) Flow cytometry phenotype characterization for donor 2. (B) Tri-lineage analysis for donor 1, demonstrating adipogenic (Oil Red O stained lipid vacuoles), osteogenic (Alizarin Red S stained mineralized matrix), and chondrogenic (Alcian Blue stained matrix) potential. (A) Flow cytometry characterization for donor 1. Flow cytometry analysis and tri-lineage differentiation of hBMSC from donors 1 and 2. The Creative Commons Public Domain Dedication waiver ( ) applies to the data made available in this article, unless otherwise stated in a credit line to the data.Īdditional file 1: Supplementary Figure 1. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder.

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