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<t>Microfluidic</t> culture of ECs to decouple pressure and flow effect. (A) Schematic of the flow circuit connecting the culture channel and a resistor channel in series with the syringe pump. Different resistor channels were designed with specific width and height to change the hydrostatic pressure P set at the outlet of culture channels. (B) Representative images of ECs cultured at shear stress τ w = 5 dyne/cm 2 show distinct cell morphology and alignment for P set = 0 (i) and 60 mmHg (ii). Red: F-actin; green: KI-67; blue: nuclei; gray: VECad. (C) Quantification of cell area, alignment angle, aspect ratio, and number of nuclei per 40x field for two pressure conditions. * P < 0.05. (D) Representative immunostaining images of ECs show different polarization of Notch1-ECD (green) at different pressure conditions under flow. (E, F, G, H) Transcriptional changes of ECs in response to flow and pressure. (E) Principal component analysis showing the separation of static and flow conditions in PC1 and mild pressure separation in PC2. (F) Venn diagram of differentially expressed gene numbers comparing the effect of flow and pressure on EC response. Red: upregulated genes; blue: downregulated genes. 5_0: τ w = 5 dyne/cm 2 and P set = 0 mmHg; and 5_60: τ w = 5 dyne/cm 2 and P set = 60 mmHg. (G) Heatmap of selected flow-responsive markers (upper) and differentially expressed cell-cycle-related genes comparing static and pressure groups. Colormap: log2(CPM) with minimum value in blue and maximum in red. (H) GO term analysis and the number of genes associated comparing high vs low pressure at the same flow shear condition.
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<t>Microfluidic</t> culture of ECs to decouple pressure and flow effect. (A) Schematic of the flow circuit connecting the culture channel and a resistor channel in series with the syringe pump. Different resistor channels were designed with specific width and height to change the hydrostatic pressure P set at the outlet of culture channels. (B) Representative images of ECs cultured at shear stress τ w = 5 dyne/cm 2 show distinct cell morphology and alignment for P set = 0 (i) and 60 mmHg (ii). Red: F-actin; green: KI-67; blue: nuclei; gray: VECad. (C) Quantification of cell area, alignment angle, aspect ratio, and number of nuclei per 40x field for two pressure conditions. * P < 0.05. (D) Representative immunostaining images of ECs show different polarization of Notch1-ECD (green) at different pressure conditions under flow. (E, F, G, H) Transcriptional changes of ECs in response to flow and pressure. (E) Principal component analysis showing the separation of static and flow conditions in PC1 and mild pressure separation in PC2. (F) Venn diagram of differentially expressed gene numbers comparing the effect of flow and pressure on EC response. Red: upregulated genes; blue: downregulated genes. 5_0: τ w = 5 dyne/cm 2 and P set = 0 mmHg; and 5_60: τ w = 5 dyne/cm 2 and P set = 60 mmHg. (G) Heatmap of selected flow-responsive markers (upper) and differentially expressed cell-cycle-related genes comparing static and pressure groups. Colormap: log2(CPM) with minimum value in blue and maximum in red. (H) GO term analysis and the number of genes associated comparing high vs low pressure at the same flow shear condition.
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Microfluidic culture of ECs to decouple pressure and flow effect. (A) Schematic of the flow circuit connecting the culture channel and a resistor channel in series with the syringe pump. Different resistor channels were designed with specific width and height to change the hydrostatic pressure P set at the outlet of culture channels. (B) Representative images of ECs cultured at shear stress τ w = 5 dyne/cm 2 show distinct cell morphology and alignment for P set = 0 (i) and 60 mmHg (ii). Red: F-actin; green: KI-67; blue: nuclei; gray: VECad. (C) Quantification of cell area, alignment angle, aspect ratio, and number of nuclei per 40x field for two pressure conditions. * P < 0.05. (D) Representative immunostaining images of ECs show different polarization of Notch1-ECD (green) at different pressure conditions under flow. (E, F, G, H) Transcriptional changes of ECs in response to flow and pressure. (E) Principal component analysis showing the separation of static and flow conditions in PC1 and mild pressure separation in PC2. (F) Venn diagram of differentially expressed gene numbers comparing the effect of flow and pressure on EC response. Red: upregulated genes; blue: downregulated genes. 5_0: τ w = 5 dyne/cm 2 and P set = 0 mmHg; and 5_60: τ w = 5 dyne/cm 2 and P set = 60 mmHg. (G) Heatmap of selected flow-responsive markers (upper) and differentially expressed cell-cycle-related genes comparing static and pressure groups. Colormap: log2(CPM) with minimum value in blue and maximum in red. (H) GO term analysis and the number of genes associated comparing high vs low pressure at the same flow shear condition.

Journal: Vascular Biology

Article Title: Under pressure: integrated endothelial cell response to hydrostatic and shear stresses

doi: 10.1530/VB-25-0015

Figure Lengend Snippet: Microfluidic culture of ECs to decouple pressure and flow effect. (A) Schematic of the flow circuit connecting the culture channel and a resistor channel in series with the syringe pump. Different resistor channels were designed with specific width and height to change the hydrostatic pressure P set at the outlet of culture channels. (B) Representative images of ECs cultured at shear stress τ w = 5 dyne/cm 2 show distinct cell morphology and alignment for P set = 0 (i) and 60 mmHg (ii). Red: F-actin; green: KI-67; blue: nuclei; gray: VECad. (C) Quantification of cell area, alignment angle, aspect ratio, and number of nuclei per 40x field for two pressure conditions. * P < 0.05. (D) Representative immunostaining images of ECs show different polarization of Notch1-ECD (green) at different pressure conditions under flow. (E, F, G, H) Transcriptional changes of ECs in response to flow and pressure. (E) Principal component analysis showing the separation of static and flow conditions in PC1 and mild pressure separation in PC2. (F) Venn diagram of differentially expressed gene numbers comparing the effect of flow and pressure on EC response. Red: upregulated genes; blue: downregulated genes. 5_0: τ w = 5 dyne/cm 2 and P set = 0 mmHg; and 5_60: τ w = 5 dyne/cm 2 and P set = 60 mmHg. (G) Heatmap of selected flow-responsive markers (upper) and differentially expressed cell-cycle-related genes comparing static and pressure groups. Colormap: log2(CPM) with minimum value in blue and maximum in red. (H) GO term analysis and the number of genes associated comparing high vs low pressure at the same flow shear condition.

Article Snippet: Syringes were connected to a microfluidic flow sensor (Elveflow MFS-80, France), a bare PDMS culture tube, a microfluidic pressure sensor (Elveflow MPS-1), and bare PDMS resistors of different lengths.

Techniques: Cell Culture, Shear, Immunostaining