Journal: Advanced Science

Article Title: Pericytes Promote More Vascularization than Stromal Cells via an Interleukin‐6‐Dependent Mechanism in Microfluidic Chips

doi: 10.1002/advs.202408131

Figure Lengend Snippet: Replication of the two main vascularization processes in microfluidic devices. A) Main mechanisms of vessel formation in the human body. B) A schematic drawing of the microfluidic chip shows the internal hydrogel channel (blue) and the surrounding parallel media channels (red). C) Fluorescence images showing freshly seeded endothelial cells (Vybrant DiD, magenta) and supporting cells (Vybrant DiO, yellow), which can be either pericytes or stromal cells, in two different set‐ups to study vasculogenesis (left) and angiogenesis (right). White arrows mark the interface between the hydrogel and media channel where the endothelial cells attach. Scale bars: 500 µm.

Article Snippet: To compare the processes of vasculogenesis and angiogenesis ( Figure 1 A ), and how pericytes and stromal cells act in both, we used a commercially available three‐channel microfluidic chip from AIM Biotech, Singapore (Figure ).

Techniques: Fluorescence

Journal: Advanced Science

Article Title: Pericytes Promote More Vascularization than Stromal Cells via an Interleukin‐6‐Dependent Mechanism in Microfluidic Chips

doi: 10.1002/advs.202408131

Figure Lengend Snippet: Comparison of the pro‐angiogenic potential of pericytes and stromal cells. A) Immunofluorescence images of the angiogenic sprouting within the microfluidic chips co‐cultured with either pericytes or stromal cells, stained for nuclei (DAPI; cyan), CD31 showing the endothelial vessel‐like structures (magenta) and the supporting cells’ F‐actin cytoskeleton (Phalloidin; yellow). F‐actin is not visible within the endothelial cells since it was subtracted from the picture during the image processing to increase clarity. Scale bar: 200 µm (left), 100 µm (right). B–E) Comparison of total vessel volume ( p = 0.0428), length ( p = 0.0340), branching points ( p = 0.0329), and average diameter ( p = 0.6508) between both co‐culture conditions (n = 3, unpaired t‐test, p < 0.05 is considered significant). F) Comparison of the distance between supporting cells and the nearest vessel‐like structure ( p < 0.0001). The trunked violin plots depict summary statistics and the kernel density estimation to show the frequency distribution of each condition. The middle line represents the median (n = 3, Mann‐Whitney test, p < 0.05 is considered significant). G) Comparison of supporting cells’ average sphericity ( p = 0.2667, n = 3, unpaired t‐test, p < 0.05 is considered significant). Pericytes and stromal cells from 3 different donors each were used for the co‐cultures (n = 3), while the endothelial cell donor remained constant. Asterisks indicate a statistical significance (* p < 0.05, **** p < 0.0001).

Article Snippet: To compare the processes of vasculogenesis and angiogenesis ( Figure 1 A ), and how pericytes and stromal cells act in both, we used a commercially available three‐channel microfluidic chip from AIM Biotech, Singapore (Figure ).

Techniques: Comparison, Immunofluorescence, Cell Culture, Staining, Co-Culture Assay, MANN-WHITNEY

Journal: Advanced Science

Article Title: Pericytes Promote More Vascularization than Stromal Cells via an Interleukin‐6‐Dependent Mechanism in Microfluidic Chips

doi: 10.1002/advs.202408131

Figure Lengend Snippet: Comparison of pericytes and stromal cells in promoting vasculogenesis of endothelial cells. A) Immunofluorescence images of the vasculogenesis process within the microfluidic chips co‐cultured with either pericytes or stromal cells, stained for nuclei (DAPI; cyan), CD31 showing the endothelial vessel‐like structures (magenta) and the supporting cells’ actin cytoskeleton (F‐actin, Phalloidin; yellow). Scale bars: 200 µm. B) Comparison of total vessel volume between both co‐culture conditions ( p = 0.0223, n = 3, unpaired t‐test, p < 0.05 is considered significant). C) Comparison of the distance between each type of supporting cell and the nearest vessel‐like structure. The trunked violin plots depict summary statistics and the kernel density estimation to show the frequency distribution of each condition. The middle line represents the median (n = 3, Mann‐Whitney test, p < 0.05 is considered significant). D) Comparison of the final total number of endothelial cells (ECs; p = 0.1372, n = 3, unpaired t‐test, p < 0.05 is considered significant) and supporting cells (SPs; p = 0.999, n = 3, Mann‐Whitney test, p < 0.05 is considered significant). E) Comparison of supporting cells’ average sphericity ( p = 0.2099, n = 3, unpaired t‐test, p < 0.05 is considered significant). F) Quantification and comparison of ten cytokines tightly related to vascularization, namely angiopoietin 1, angiopoietin 2, vascular endothelial growth factor (VEGF), epidermal growth factor (EGF), fibroblast growth factor 2 (FGF‐2), interleukin‐6 (IL‐6) and interleukin‐8 (IL‐8), tumor necrosis factor (TNF), PECAM‐1 and placenta growth factor (PlGF). Statistical values can be found in Table (Supporting Information) (n = 3, p < 0.05 is considered significant). Pericytes and stromal cells from 3 different donors each were used for the co‐cultures (n = 3), while the endothelial cell donor remained constant. Asterisks indicate statistical significance (* p < 0.05, ** p < 0.01, **** p < 0.0001).

Article Snippet: To compare the processes of vasculogenesis and angiogenesis ( Figure 1 A ), and how pericytes and stromal cells act in both, we used a commercially available three‐channel microfluidic chip from AIM Biotech, Singapore (Figure ).

Techniques: Comparison, Immunofluorescence, Cell Culture, Staining, Co-Culture Assay, MANN-WHITNEY