normal human bronchial smooth muscle cells  (Lonza)

Journal: Advanced healthcare materials

Article Title: Stiffness Mediated-Mechanosensation of Airway Smooth Muscle Cells on Linear Stiffness Gradient Hydrogels.

doi: 10.1002/adhm.202304254

Figure Lengend Snippet: Figure 1. Measurement of ex vivo airway smooth muscle (ASM) stiffness. A) A custom 3D-printed platform was developed to accommodate the dimen- sions of porcine tracheal ASM strips (40 mm in length × 10 mm in width) and facilitate longitudinal stretch at 1 mm increments. B) The 3D-printed platform was mounted onto a motorized linear translation stage for uniaxial compression of the ASM strip. C) Stiffness of ASM from proximal and distal locations of the porcine trachea (proximal N = 6, distal N = 6) were assessed at 0%, 5%, and 10% levels and under 10% and 40% uniaxial compressions. Data are presented as mean ± SD.

Article Snippet: Cell Culture: Primary human bronchial and tracheal smooth muscle cells (PCS-130-011; ATCC) were cultured in growth media containing DMEM (Life Technologies) with 5% FBS (Life Technologies) and 1 × antibiotic-antimycotic (Thermo Scientific).

Techniques: Ex Vivo, Stripping Membranes

Journal: Advanced healthcare materials

Article Title: Stiffness Mediated-Mechanosensation of Airway Smooth Muscle Cells on Linear Stiffness Gradient Hydrogels.

doi: 10.1002/adhm.202304254

Figure Lengend Snippet: Figure 3. Profiling mechanosensitivity of human airway smooth muscle cell (hASMC) morphology to extracellular matrix (ECM) stiffness and protein composition. A) hASMC were cultured on linear stiffness gradient hydrogels (Figure 2) with either collagen I (ColI), fibronectin (Fn), or laminin (Ln) coating for 96 h before being fixed, stained, and imaged at eight different points of stiffness. Identification of cells by F-actin and DAPI staining was automated using CellProfiler. B) Cell size was positively correlated with stiffness (Pearson’s correlation: ColI R2 = 0.86, Fn R2 = 0.84, Ln R2 = 0.67). No significant differences between the slopes of ECM proteins were observed (Figure S4A, Supporting Information), however, significant differences were observed between the intercepts of the ECM proteins (linear regression: p < 0.0001) (Figure S4D, Supporting Information). C) Nuclear size was positively correlated with stiffness (Pearson’s correlation: ColI R2 = 0.87, Fn R2 = 0.80, Ln R2 = 0.77). Borderline significance in the differences between ECM protein slopes was found with linear regression (linear regression: p = 0.05). No correlations with stiffness were observed for D) cell aspect ratio and E) cell form factor. F) Nuclear aspect ratio showed no correlation with the stiffness however, G) nuclear form factor showed an inverse forrelation with stiffness on ColI (Pearson’s correlation: R2 = 0.55), but none with F) nuclear aspect ratio. Further comparisons for differences between data for all three ECM proteins at each stiffness (for all morphological assessments) were conducted with one-way ANOVA (Table S1, Supporting Information). Simple linear regression lines are plotted on graphs with 95% confidence intervals and shades accordingly. Data on graphs are presented as mean ± SEM. For (B–(G), N = 6 repeats, n > 4500 cells analyzed. *On graphs indicate a significant correlation with stiffness, p < 0.05.

Article Snippet: Cell Culture: Primary human bronchial and tracheal smooth muscle cells (PCS-130-011; ATCC) were cultured in growth media containing DMEM (Life Technologies) with 5% FBS (Life Technologies) and 1 × antibiotic-antimycotic (Thermo Scientific).

Techniques: Cell Culture, Staining

Journal: Advanced healthcare materials

Article Title: Stiffness Mediated-Mechanosensation of Airway Smooth Muscle Cells on Linear Stiffness Gradient Hydrogels.

doi: 10.1002/adhm.202304254

Figure Lengend Snippet: Figure 5. Profiling human airway smooth muscle cell (hASMC) morphology, contractile function, and mechanomarker expression with inhibition of mechanosensation. A) hASMC were cultured on linear stiffness gradient hydrogels coated with fibronectin (Fn) as control (data from Figures 3 and 4) and treated with blebbistatin and Y27632 for 96 h before being fixed, stained and imaged at eight points of stiffness. B) Cell size with both blebbistatin and Y27632 were attenuated with controls. Only cells inhibited with blebbistatin exhibited a positive correlation with stiffness (Pearson’s correlation: R2

Article Snippet: Cell Culture: Primary human bronchial and tracheal smooth muscle cells (PCS-130-011; ATCC) were cultured in growth media containing DMEM (Life Technologies) with 5% FBS (Life Technologies) and 1 × antibiotic-antimycotic (Thermo Scientific).

Techniques: Expressing, Inhibition, Cell Culture, Control, Staining