transwell permeable support membrane coated with matrigel  (Corning Life Sciences)

Journal: Cells

Article Title: Nanospheres Loaded with Curcumin Improve the Bioactivity of Umbilical Cord Blood-Mesenchymal Stem Cells via c-Src Activation during the Skin Wound Healing Process

doi: 10.3390/cells9061467

Figure Lengend Snippet: The effect of CN on migration of human UCB-MSCs. ( A , B ) Dose and time responses of Curcumin (10 ng/mL and 1 µg/mL) and CN (100 pg/mL and 10 ng/mL) in the migration of UCB-MSCs were quantified and visualized with the confocal microscopy. Scale bar represents 100 μm (magnification, × 100). * p ≤ 0.01 versus control. ( C ) Lamellipodial extrusions in cells treated with CN of 100 pg/mL for 60 min are shown. Cells were fixed and labeled with phalloidin-Alexa Fluor 488 (green) to identify the leading edge of lamellipodia (arrows). Dashed lines indicate the leading edges. n = 3. Scale bars represent 100 μm (magnification, × 400). ( D ) The migration capacity of UCB-MSCs treated with CN for 24 h was assessed in transwell permeable support with 8.0 μm pore size membrane coated with Matrigel. Data represent the means ± S.E. n = 3. Scale bars represent 100 μm (magnification, × 200). * p ≤ 0.001 versus control. ( E ) The proliferation of UCB-MSCs treated with CN for 24 h was determined by EZ-CYTOX cell proliferation kit. Data represent the means ± S.E. n = 3.

Article Snippet: An in vitro transwell migration assay was conducted using a Transwell permeable support with 8.0 µm pore size membrane coated with Matrigel (BD Biosciences) according to the manufacturer’s instruction.

Techniques: Migration, Confocal Microscopy, Labeling

Journal: Cells

Article Title: Nanospheres Loaded with Curcumin Improve the Bioactivity of Umbilical Cord Blood-Mesenchymal Stem Cells via c-Src Activation during the Skin Wound Healing Process

doi: 10.3390/cells9061467

Figure Lengend Snippet: CN regulates the activation of c-Src and PKC. ( A ) Time responses of phosphorylation of c-Src in cells treated with CN are shown. Data represent the means ± S.E. n = 3. * p ≤ 0.001 vs. 0 min. ( B ) Membrane translocation of p-c-Src (green) was determined by confocal microscopy. Propidium iodide (PI) was used for nuclear counterstaining (red). n = 3. Scale bars represent 100 μm (magnification, × 400). ( C ) Cells were pretreated with c-Src inhibitor, PP2 (10 µM) for 30 min prior exposure to CN for 24 h. Cell migration was determined by wound-healing migration assay (left panel) and quantified (right panel). Scale bar represents 100 µm (magnification, × 100). n = 3. * p ≤ 0.001 vs. control. # p ≤ 0.05 vs. CN alone. ( D ) The migration capacity of UCB-MSCs treated with CN for 24 h was assessed in transwell permeable support with 8.0 μm pore size membrane coated with Matrigel. Data represent the means ± S.E. n = 3. Scale bars represent 100 μm (magnification, × 200). * p ≤ 0.01 vs. control. # p ≤ 0.01 vs. CN alone. ( E ) Time responses of phosphorylation of p-PKC in cells treated with CN are shown. Data represent the means ± S.E. n = 3. * p ≤ 0.001 vs. control. ( F ) Cells were pretreated with PP2 for 30 min prior exposure to CN for 60 min. The level of phosphorylation of PKC was determined by western blot. Data represent the means ± S.E. n = 3. * p ≤ 0.05 vs. control. # p ≤ 0.01 vs. CN alone. ( G ) Membrane translocation of p-PKC (green) was determined by confocal microscopy. Propidium iodide (PI) was used for nuclear counterstaining (red). n = 3. Scale bars represent 100 μm (magnification, × 400). ( H ) Cells were pretreated with PKC inhibitor, Bisindolylmaleimide I (Bis I, 10 µM) for 30 min prior exposure to CN for 24 h. Cell migration was determined by wound-healing migration assay (left panel) and quantified (right panel). Scale bar represents 100 µm (magnification, × 100). n = 3. * p ≤ 0.001 vs. control. # p ≤ 0.01 vs. CN alone. ( I ) The migration capacity of UCB-MSCs treated with CN for 24 h was assessed in transwell permeable support with 8.0 μm pore size membrane coated with Matrigel. Data represent the means ± S.E. n = 3. Scale bars represent 100 μm (magnification, × 200). * p ≤ 0.01 vs. control. # p ≤ 0.05 vs. CN alone. ( A , E , F ) ROD is the abbreviation for relative optical density.

Article Snippet: An in vitro transwell migration assay was conducted using a Transwell permeable support with 8.0 µm pore size membrane coated with Matrigel (BD Biosciences) according to the manufacturer’s instruction.

Techniques: Activation Assay, Translocation Assay, Confocal Microscopy, Migration, Western Blot

Journal: Cells

Article Title: Nanospheres Loaded with Curcumin Improve the Bioactivity of Umbilical Cord Blood-Mesenchymal Stem Cells via c-Src Activation during the Skin Wound Healing Process

doi: 10.3390/cells9061467

Figure Lengend Snippet: CN uniquely regulates the ERK pathway responsible for the cell migration. ( A ) Time responses of phosphorylation of mitogen-activated protein kinases (MAPK) in cells exposed to CN for 120 min are shown. Data represent the means ± S.E. n = 3. * p ≤ 0.01 vs. 0 min. ( B ) Cells were pretreated with Bis I for 30 min prior exposure to CN for 60 min. The level of phosphorylation of MAPK was determined by western blot. Data represent the means ± S.E. n = 4. * p ≤ 0.01 vs. control. # p ≤ 0.05 vs. CN alone. ( C ) Cells were pretreated with ERK inhibitor, PD98059 (10 µM) for 30 min prior exposure to CN for 24 h. Cell migration was determined by wound-healing migration assay (left panel) and quantified (right panel). Scale bar represents 100 µm (magnification, × 100). n = 3. * p ≤ 0.01 vs. control. # p ≤ 0.001 vs. CN alone. ( D ) The migration capacity of UCB-MSCs treated with CN for 24 h was assessed in transwell permeable support with 8.0 μm pore size membrane coated with Matrigel. Data represent the means ± S.E. n = 3. Scale bars represent 100 μm (magnification, × 200). * p ≤ 0.001 vs. control. # p ≤ 0.01 vs. CN alone. ( A , B ) ROD is the abbreviation for relative optical density.

Article Snippet: An in vitro transwell migration assay was conducted using a Transwell permeable support with 8.0 µm pore size membrane coated with Matrigel (BD Biosciences) according to the manufacturer’s instruction.

Techniques: Migration, Western Blot

Journal: Cells

Article Title: Nanospheres Loaded with Curcumin Improve the Bioactivity of Umbilical Cord Blood-Mesenchymal Stem Cells via c-Src Activation during the Skin Wound Healing Process

doi: 10.3390/cells9061467

Figure Lengend Snippet: Regulatory effect of CN on the activation of NF-κB and the cytoskeletal reorganization. ( A ) Time responses of phosphorylation of IκBα and NF-κB in cell treated with CN are shown. Phosphorylation of IκBα and NF-κB were determined by western blot. Data represent means ± S.E. n = 3. * p ≤ 0.01 vs. 0 h. ( B ) Cells were pretreated with PD98059 for 30 min prior exposure to CN for 2 h. The level of phosphorylation of NF-κB was determined by western blot. Data represent the means ± S.E. n = 4. * p ≤ 0.01 vs. control. # p ≤ 0.05 vs. CN alone. ( C ) The increased level of p-NF-κB (green) was determined by confocal microscopy. Propidium iodide (PI) was used for nuclear counterstaining (red). n = 3. Scale bars represent 100 μm (magnification, × 400). ( D ) Cells were pretreated with NF-κB inhibitor, Bay 11-7082 (10 µM) for 30 min prior exposure to CN for 24 h. Cell migration was determined by wound-healing migration assay (left panel) and quantified (right panel). Scale bar represents 100 µm (magnification, × 100). n = 3. * p ≤ 0.01 vs. control. # p ≤ 0.05 vs. CN alone. ( E ) The migration capacity of UCB-MSCs treated with CN for 24 h was assessed in transwell permeable support with 8.0-μm pore size membrane coated with Matrigel. Data represent the means ± S.E. n = 3. Scale bars represent 100 μm (magnification, × 200). * p ≤ 0.01 vs. control. # p ≤ 0.05 vs. CN alone. ( F ) Time responses of CN in the expression of α-actinin, PFN-1, and F-actin are shown (0–24 h). Data represent means ± S.E. n = 3. * p ≤ 0.01 vs. 0 h. The expression of α-actinin (green) ( G ) and PFN-1(green) ( H ) was determined by confocal microscopy. PI was used for nuclear counterstaining (red). n = 3. Scale bars represent 100 μm (magnification, × 400). ( I ) Cells were pretreated with Bay 11-7082 for 30 min prior exposure to CN for 24 h. Expression of α-actinin, F-actin, and PFN-1 were determined by western blot. Data represent the means ± S.E. n = 4. * p ≤ 0.01 vs. control. # p ≤ 0.05 vs. CN alone. ( A , B , F , I ) ROD is the abbreviation for relative optical density.

Article Snippet: An in vitro transwell migration assay was conducted using a Transwell permeable support with 8.0 µm pore size membrane coated with Matrigel (BD Biosciences) according to the manufacturer’s instruction.

Techniques: Activation Assay, Western Blot, Confocal Microscopy, Migration, Expressing