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Spatial patterning of endothelial cells and vascular network formation using ultrasound standing wave fields
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10.1121/1.4812867
/content/asa/journal/jasa/134/2/10.1121/1.4812867
http://aip.metastore.ingenta.com/content/asa/journal/jasa/134/2/10.1121/1.4812867
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Figures

Image of FIG. 1.
FIG. 1.

Spatial patterning of cells within collagen hydrogels using ultrasound standing wave fields. Unpolymerized solutions of type I collagen and endothelial cells (4 × 10 cell/ml) were exposed for 15 min during the polymerization process to either a 1- or 2-MHz CW ultrasound standing wave field. Samples were exposed to various peak positive pressure amplitudes (measured at standing wave field pressure antinodes) and calculated values. Resultant collagen gels were analyzed for cell location using phase-contrast microscopy. Representative side-view images of cell-embedded collagen gels are shown for 1-MHz (A) and 2-MHz (B) experiments ( = 3). Scale bar, 200 m.

Image of FIG. 2.
FIG. 2.

Dependence of cell patterns on temporal average intensity. Unpolymerized solutions of type I collagen and endothelial cells (4 × 10 cell/ml) were exposed during polymerization to a 1-MHz ultrasound standing wave field. Samples were exposed using (A) a constant ultrasound standing wave field peak positive pressure amplitude (measured at a pressure antinode) of 0.2 MPa and various values, or (B) a constant of 0.28 W/cm and various ultrasound standing wave field peak positive pressure amplitudes. Resultant collagen gels were analyzed for cell location using phase-contrast microscopy. Representative side-view images of cell-embedded collagen gels are shown for both the constant pressure (A) and constant (B) experiments ( = 3). Scale bar, 200 m.

Image of FIG. 3.
FIG. 3.

Morphology of vascular networks produced using ultrasound standing wave fields at 1 MHz. Unpolymerized solutions of type I collagen and endothelial cells (1 × 10 cell/ml) were exposed during polymerization to a 1-MHz CW ultrasound standing wave field. Samples were exposed using antinode pressure amplitudes of 0, 0.1, or 0.3 MPa, which correspond to values of 0, 0.28, and 2.4 W/cm, respectively. (A) Resultant collagen gels were incubated at 37 °C for either 4 or 10 days and then analyzed using phase-contrast microscopy and MTT staining. Representative top view images are shown ( = 3). Scale bar, 100 m.

Image of FIG. 4.
FIG. 4.

Analysis of endothelial network morphology formed in response to 1-MHz ultrasound. Unpolymerized solutions of type I collagen and endothelial cells were exposed to a 1-MHz CW ultrasound standing wave field, as described in the legend to Fig. 3 . Following a 10-day incubation, samples were processed for immunofluorescence microscopy. CD31 was visualized by staining with a mouse anti-human CD31 monoclonal antibody (red). Cell nuclei were visualized by co-staining with DAPI (blue). Multiphoton microscopy was used to collect top view images along the axis in 1 m slices. Images were then projected onto the -plane using ImageJ software (NIH, Bethesda, MD). Representative -stack images are shown before [(A) and (B)] and after [(C) and (D)] image processing ( = 3). Scale bar, 50 m.

Image of FIG. 5.
FIG. 5.

Morphology of vascular networks produced using ultrasound standing wave fields at 2 MHz. Unpolymerized solutions of type I collagen and endothelial cells (1 × 10 cell/ml) were exposed during polymerization to a 2-MHz CW ultrasound standing wave field. Samples were exposed using antinode pressure amplitudes of 0, 0.08, or 0.2 MPa, corresponding to values of 0, 0.2, and 1.6 W/cm, respectively. Resultant collagen gels were incubated at 37 °C for 4 or 10 days and then analyzed using phase-contrast microscopy and MTT staining. Representative top view images are shown ( = 3). Scale bar, 100 m.

Image of FIG. 6.
FIG. 6.

Analysis of endothelial network morphology formed in response to 2-MHz ultrasound. Unpolymerized solutions of type I collagen and endothelial cells were exposed to a 2-MHz CW ultrasound standing wave field, as described in the legend to Fig. 5 . Following a 10-day incubation, samples were processed for immunofluorescence microscopy. CD31 was visualized by staining with a mouse anti-human CD31 monoclonal antibody (red). Cell nuclei were visualized by co-staining with DAPI (blue). Multiphoton microscopy was used to collect top view images along the axis in 1 m slices. Images were then projected onto the -plane using ImageJ (NIH). Representative -stack images are shown before [(A) and (B)] and after [(C) and (D)] image processing ( = 3). Scale bar, 50 m.

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/content/asa/journal/jasa/134/2/10.1121/1.4812867
2013-08-01
2014-04-23
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752b84549af89a08dbdd7fdb8b9568b5 journal.articlezxybnytfddd
Scitation: Spatial patterning of endothelial cells and vascular network formation using ultrasound standing wave fields
http://aip.metastore.ingenta.com/content/asa/journal/jasa/134/2/10.1121/1.4812867
10.1121/1.4812867
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