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

Branch Points SD

TCPComplete EC Media + VEGF 104.2 13.4

Basal EC Media 106.7 15.7Proliferation Media 63.3 11.8

Membrane

Complete EC Media + VEGF 128.8 8.7Basal EC Media 130.7 13.0

Proliferation Media 41.3 9.5

Table S2. Total Tube LengthTotal Tube

Length (microns) SD

TCPComplete EC Media + VEGF 58553 10987

Basal EC Media 52433 3466Proliferation Media 38317 8562

Membrane

Complete EC Media + VEGF 77395 8152Basal EC Media 71518 5965

Proliferation Media 18434 2058

Membranes Promote differentiation of ADSCsTR Gaborski

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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 ADSCsTR Gaborski

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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 ADSCsTR Gaborski

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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 ADSCsTR Gaborski

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