SUPPLEMENTAL INFORMATION
ADSCs were cultured in the following media conditions on either 3.0 μm pore track-etched
membranes or tissue culture treated plastic (TCP) 24-well plates. Complete EC Media + 50
ng/ml VEGF and Basal EC Media were both significantly greater than Proliferation Media
on TCP and Membranes for both Total Branch Points and for Total Tube Length.
Table S1. Total Branch Points
TCP
Membrane
Complete EC Media + VEGF
Basal EC Media
Proliferation Media
Complete EC Media + VEGF
Basal EC Media
Proliferation Media
Branch
Points
104.2
106.7
63.3
128.8
130.7
41.3
SD
13.4
15.7
11.8
8.7
13.0
9.5
Total Tube
Length
(microns)
58553
52433
38317
77395
71518
18434
SD
10987
3466
8562
8152
5965
2058
Table S2. Total Tube Length
TCP
Membrane
Complete EC Media + VEGF
Basal EC Media
Proliferation Media
Complete EC Media + VEGF
Basal EC Media
Proliferation Media
Membranes Promote differentiation of ADSCs
TR Gaborski
1
FIGURE S1. ADSC expression of CD31 after culture on tissue culture treated plastic in
complete and basal EC media. Wells of a 24-well plate were pre-coated with a 1%
Geltrex™ solution. ADSC were cultured for 6 days prior to fixation and staining. (A) ADSCs
cultured in complete EC media with 50 ng/mL of VEGF expressed a relatively uniform
distribution of CD31 that is similar to cells grown on membranes (Figure 2a). (B) ADSCs
cultured in basal EC media without growth factors expressed minimal CD31, similarly to
cells grown on membranes (Figure 2b). Images were collected with the same exposure
settings and color combined with the identical min and max fluorescence levels.
Membranes Promote differentiation of ADSCs
TR Gaborski
2
FIGURE S2. ADSC expression of pericyte markers αSMA and NG2 after culture in
various media. (A) ADSCs cultured on Geltrex treated tissue culture plastic in MSC
proliferation media showed low levels of staining for αSMA on all cells. Some cells stained
for low levels of NG2. (B) ADSCs cultured for 6 days in basal EC media on 0.5 micron pore
size membranes showed a greater number of cells with NG2 staining, but at relatively low
levels. (C) ADSCs cultured for 6 days in complete EC media with 50 ng/mL VEGF on 0.5
micron pore size membranes showed significant NG2 staining across the majority of cells.
NG2 fluorescence levels were between 2-3x greater than those in (A) and (B). All images
were collected with the same exposure settings and color combined with the identical min
and max fluorescence levels.
Membranes Promote differentiation of ADSCs
TR Gaborski
3
FIGURE S3. Representative image reconstructions of the 96-well plate angiogenesis
assays. In order to count all branch points and total tube length in each well of the
angiogenesis assays, multiple 4x bright field images were stitched together using Adobe
Photoshop. Representative images are shown of ADSC differentiated in the following
conditions prior to the angiogenesis assay: complete EC media with 50 ng/mL VEGF on
membranes (A), basal EC media on membranes (B), proliferation media on membranes (C),
complete EC media with 50 ng/mL VEGF on TCP (D), basal EC media on TCP (E) and
proliferation media on TCP (F).
Membranes Promote differentiation of ADSCs
TR Gaborski
4