Figure 1

Identification of expanded human MSCs and adenovirus transfection efficiency. (A) Representative histogram overlays of FACS analysis showing the MSC antigen profile. The blue peak represents the specific PE-labeled antibodies: CD34, CD45, CD19, CD44, CD90, and CD105. The red peak represents the corresponding isotope antibodies. (B) Positive expression of CD34, CD45, CD19, CD44, CD90, and CD105 in five independent samples was expressed as mean ± SD as evaluated by FACS. (C) Immunostaining of CD34, CD45, CD19, CD44, CD90, and CD105 in one representative sample as observed under a confocal microscope. Negative controls were performed by using the corresponding isotope antibodies. Red, PE-labeled antibodies; blue, DAPI. (D) Representative photographs showing the in vitro differentiation of MSCs into the osteogenic, adipogenic, and chondrogenic lineages. Undifferentiated MSCs were dyed with hematoxylin and eosin. (E) Representative images depicting the transfection of MSCs with 500 MOI Adv-GFP. Forty-eight hours after transfection, MSCs carrying GFP were observed under bright field (upper panel) and fluorescent field (lower panel). (F) Transfection efficiencies at different Adv-GFP titers after 48 hours in three independent MSC samples were expressed as mean ± SD as evaluated by FACS. (G) Electron micrographs showing viral particles in MSCs. MSCs were infected with 500 MOI Adv-Stat3(-). Seventy-two hours later, MSCs were collected and prepared to be observed under an electron microscope. Adenovirus particles in the MSCs were shown using the black arrow. C, scale bar = 5 μm; D, scale bar = 10 μm; E, scale bar = 10 μm; G, left scale bar = 2 μm, right scale bar = 200 nm).