fibronectin adsorption  (Cytoskeleton Inc)

Journal: ACS nano

Article Title: Stem Cell Expansion and Fate Decision on Liquid Substrates Are Regulated by Self-Assembled Nanosheets.

doi: 10.1021/acsnano.8b03865

Figure Lengend Snippet: Figure 3. Cell spreading at liquid−liquid interfaces is mediated by integrin adhesion and regulated by acto-myosin contractility. (A) Human primary keratinocyte (HPK) spreading (after 24 h) on TPS and PLL−fibronectin-functionalized oil interfaces (functionalized with PLL at pH 7.4 or 10.5, then fibronectin at neutral pH). B. Corresponding fluorescence images (Red, actin; Blue, DAPI). C. HaCaT cells spreading on BSA interfaces (TPS and Novec 7500 + PFBC) is modulated by the action of acto-myosin inhibitors (myosin inhibitor blebbistatin, 10 μM; ROCK inhibitor Y27632, 10 μM; actin polymerization inhibitor cytochalasin D, 1 μM). Cell areas determined from actin stainings (phalloidin). (D) Blocking of β1 integrins in HPK cells spreading on PLL−FN interfaces (TPS and Novec 7500 + PFBC; blocking with mouse anti-β1 integrin antibody P5D2, 1:50, 20 μg/mL). PLL deposition was carried out in pH 10.5 PBS at the surface of fluorinated oil. Cell areas determined from actin stainings (phalloidin). (E) Corresponding fluorescence microscopy images (blue, DAPI; red, phalloidin). (F) Confocal microscopy images of HPKs spreading (after 24 h) on TPS and PLL-functionalized oil interfaces. Zoom-ins correspond to the dotted boxes. (G) SICM imaging in hopping mode of HPKs spreading (after 24 h) on TPS and PLL-functionalized oil interfaces. Zoom-ins correspond to the dotted boxes. Error bars are SEM; n = 3.

Article Snippet: Additional materials and methods, additional nanosheets mechanics and PLL adsorption kinetics, fibronectin adsorption quantification, additional cell adhesion quantification and microscopy (spreading, FA formation, actin cytoskeleton assembly, integrin blocking), imaging of keratinocytes and MSCs spreading around droplets and MSCs transferred from droplets, statistical analysis reports (PDF) AUTHOR INFORMATION Corresponding Author *E-mail: j.gautrot@qmul.ac.uk.

Techniques: Blocking Assay, Microscopy, Confocal Microscopy, Imaging

Journal: ACS nano

Article Title: Stem Cell Expansion and Fate Decision on Liquid Substrates Are Regulated by Self-Assembled Nanosheets.

doi: 10.1021/acsnano.8b03865

Figure Lengend Snippet: Figure 4. Stem cell differentiation and expansion at liquid−liquid interfaces. (A) HPK differentiation on solid (TPS functionalized with fibronectin, Fn, or PLL−PEG) and liquid interfaces (Novec 7500 + 0.01 mg/mL PFBC; oil-Fn-pH7, PLL at pH 7.4 then Fn; oil-Fn-pH10, PLL at pH 10.5 then Fn; oil-PEG-pH10, PLL−PEG at pH 10.5) in differentiation medium (FAD) and basal medium (KSFM). Error bars are SEM; n = 3. (B) Fluorescence microscopy images (red, actin; green, involucrin; blue, DAPI) corresponding to some of these conditions. (C) MSC proliferation profile on fluorinated oil interfaces functionalized with PLL−fibronectin (Fn) at pH 10.5 (PLL, 100 μg/mL; Fn, 10 μg/ mL; red, TPS; blue, Novec 7500 + 0.00125 mg/mL PFBC). Error bars are SEM; n = 3. Images are corresponding nuclear stainings at day 5 (Hoechst, scale bars are 200 μm). (D) Confocal microscopy images of MSCs spreading (after 24 h) on TPS and PLL-functionalized oil. Zoom-ins correspond to the dotted boxes. (E) Micrographs of MSCs cultured on emulsions for 7 days in growth medium (top, bright field; middle and bottom, epifluorescence images of Hoechst stainings).

Article Snippet: Additional materials and methods, additional nanosheets mechanics and PLL adsorption kinetics, fibronectin adsorption quantification, additional cell adhesion quantification and microscopy (spreading, FA formation, actin cytoskeleton assembly, integrin blocking), imaging of keratinocytes and MSCs spreading around droplets and MSCs transferred from droplets, statistical analysis reports (PDF) AUTHOR INFORMATION Corresponding Author *E-mail: j.gautrot@qmul.ac.uk.

Techniques: Fluorescence, Microscopy, Confocal Microscopy, Cell Culture

Journal: Soft matter

Article Title: Microgel Film Dynamics Modulate Cell Adhesion Behavior

doi: 10.1039/C3SM52518J

Figure Lengend Snippet: Fluorescence intensity values for fluorescent fibronectin adsorbed into microgel films at 37°C. Error bars are standard deviations. Results are statistically significant with all p < 0.0045 (n=3).

Article Snippet: Protein Adsorption Assay Human plasma fibronectin (FN, Mw: 400k – 500k, 500 μg/mL, Invitrogen) was coupled with Alexa Fluor ® 488 succinimidyl ester (1 mg/mL, Molecular Probes) in 0.1M NaHCO 3 (pH 9.0, 100 μL) for 2 hours at room temperature.

Techniques: Fluorescence