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    Becton Dickinson matrigel
    Effects of Wnt inhibitors and recombinant Wnt ligands. Shown are representative fluorescent images of BAR expression in ELF1-BAR hESCs cultured on <t>Matrigel</t> and ( A ) treated as in or ( B ( n = 3 biological replicates). (Scale
    Matrigel, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/matrigel/product/Becton Dickinson
    Average 86 stars, based on 1 article reviews
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    86
    Corning Life Sciences matrigel
    Further examples for the strain stiffening effect. Single cells were seeded on <t>Matrigel</t> and the stiffness of the gel between the cells was measured with an AFM. As control, substrate stiffness in a cell-free area was also measured. The pseudo-coloured stiffness map shows the relative Young’s modulus normalized to the control area. From the overlay of the microscope Image and the stiffness map it can be seen that the cells formed a stiff bridge between them even before cellular protrusions form. In a time range from 30 min to 2 h, the cells used the stiffness line for locating each other and for contacting. Scale bar: 20 µm.
    Matrigel, supplied by Corning Life Sciences, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/matrigel/product/Corning Life Sciences
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    matrigel - by Bioz Stars, 2021-03
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    86
    Becton Dickinson growth factor reduced matrigel
    LOXL2 expression induces ErbB2 activation through production of 2 O 2 . (A)Production of H 2 O 2 on <t>Matrigel</t> was measured as an activity read-out in the 10A cont and 10A L2 cells, by using a commercially available kit and following the manufacturer's instructions (Abcam). 10A L2 cells produced higher levels of H 2 O 2 , as expected, because of LOXL2 overexpression. (B) The 200 U/ml catalase (+cat) was added to remove H 2 O 2 produced by LOXL2. Western blotting revealed that pErbB2 was decreased in 10A L2 cells treated with catalase (L2+cat) in comparison with untreated cells (L2). (C) Addition of H 2 O 2 to 10A cont cells strongly induced ErbB2 activation. Taken together, these results indicate that, by removing H 2 O 2 , we can significantly abrogate ErbB2 activation in 10A L2 cells, and addition of H 2 O 2 activated ErbB2.
    Growth Factor Reduced Matrigel, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/growth factor reduced matrigel/product/Becton Dickinson
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    growth factor reduced matrigel - by Bioz Stars, 2021-03
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    86
    Becton Dickinson matrigel basement membrane matrix
    The E-cadherin expression of EpH4 cells 7 days after incubation of EpH4 cells with MM, GM, and <t>matrigel-coated</t> GM or without microspheres and EpH4 cells were cultured by the conventional 3D and 2D monolayer methods in the absence (□) or presence of PI3K inhibitor LY294002 (■). The number of EpH4 cells added initially was 1 × 10 4 /well while that of microspheres was 1 × 10 2 or 1 × 10 3 /well. *, p
    Matrigel Basement Membrane Matrix, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/matrigel basement membrane matrix/product/Becton Dickinson
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    matrigel basement membrane matrix - by Bioz Stars, 2021-03
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    Image Search Results


    Effects of Wnt inhibitors and recombinant Wnt ligands. Shown are representative fluorescent images of BAR expression in ELF1-BAR hESCs cultured on Matrigel and ( A ) treated as in or ( B ( n = 3 biological replicates). (Scale

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Wnt/β-catenin signaling promotes self-renewal and inhibits the primed state transition in naïve human embryonic stem cells

    doi: 10.1073/pnas.1613849113

    Figure Lengend Snippet: Effects of Wnt inhibitors and recombinant Wnt ligands. Shown are representative fluorescent images of BAR expression in ELF1-BAR hESCs cultured on Matrigel and ( A ) treated as in or ( B ( n = 3 biological replicates). (Scale

    Article Snippet: In some experiments, where indicated cells were grown on Matrigel (coating concentration 0.035 g/cm2 , BD #354234) in MEF-CM with the supplements described above.

    Techniques: Recombinant, Expressing, Cell Culture

    ( A ) Western blot of AXIN1 after naïve ELF1 hESCs were treated with the indicated small molecules and cultured on Matrigel for 3 d. HSP90 was used as loading control. ( B ) Representative bright field (BF) and fluorescent (Venus) images of naïve

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Wnt/β-catenin signaling promotes self-renewal and inhibits the primed state transition in naïve human embryonic stem cells

    doi: 10.1073/pnas.1613849113

    Figure Lengend Snippet: ( A ) Western blot of AXIN1 after naïve ELF1 hESCs were treated with the indicated small molecules and cultured on Matrigel for 3 d. HSP90 was used as loading control. ( B ) Representative bright field (BF) and fluorescent (Venus) images of naïve

    Article Snippet: In some experiments, where indicated cells were grown on Matrigel (coating concentration 0.035 g/cm2 , BD #354234) in MEF-CM with the supplements described above.

    Techniques: Western Blot, Cell Culture

    Wnt/β-catenin signaling is dispensable for maintaining pluoripotency in H1-4iLIF hESCs. ( A ) Representative plots of flow cytometry for Tra1-60 and CD9 in H1-4iLIF hESCs grown on Matrigel and with the indicated small molecules for 3 d (mean ±

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Wnt/β-catenin signaling promotes self-renewal and inhibits the primed state transition in naïve human embryonic stem cells

    doi: 10.1073/pnas.1613849113

    Figure Lengend Snippet: Wnt/β-catenin signaling is dispensable for maintaining pluoripotency in H1-4iLIF hESCs. ( A ) Representative plots of flow cytometry for Tra1-60 and CD9 in H1-4iLIF hESCs grown on Matrigel and with the indicated small molecules for 3 d (mean ±

    Article Snippet: In some experiments, where indicated cells were grown on Matrigel (coating concentration 0.035 g/cm2 , BD #354234) in MEF-CM with the supplements described above.

    Techniques: Flow Cytometry, Cytometry

    Wnt/β-catenin signaling is dispensable for maintaining pluripotency in ELF1 hESCs. ( A ) Representative plots of flow cytometry for Tra1-60 and CD9 in ELF1 hESCs grown on Matrigel and with the indicated small molecules for 3 d or after transfection

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Wnt/β-catenin signaling promotes self-renewal and inhibits the primed state transition in naïve human embryonic stem cells

    doi: 10.1073/pnas.1613849113

    Figure Lengend Snippet: Wnt/β-catenin signaling is dispensable for maintaining pluripotency in ELF1 hESCs. ( A ) Representative plots of flow cytometry for Tra1-60 and CD9 in ELF1 hESCs grown on Matrigel and with the indicated small molecules for 3 d or after transfection

    Article Snippet: In some experiments, where indicated cells were grown on Matrigel (coating concentration 0.035 g/cm2 , BD #354234) in MEF-CM with the supplements described above.

    Techniques: Flow Cytometry, Cytometry, Transfection

    Wnt inhibition does not induce apoptosis or affect specific stages of the cell cycle in naïve ELF1 hESCs. ( A ) Representative flow cytometry histogram of the PARP+ population in naïve ELF1 hESCs grown on Matrigel and with the indicated

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Wnt/β-catenin signaling promotes self-renewal and inhibits the primed state transition in naïve human embryonic stem cells

    doi: 10.1073/pnas.1613849113

    Figure Lengend Snippet: Wnt inhibition does not induce apoptosis or affect specific stages of the cell cycle in naïve ELF1 hESCs. ( A ) Representative flow cytometry histogram of the PARP+ population in naïve ELF1 hESCs grown on Matrigel and with the indicated

    Article Snippet: In some experiments, where indicated cells were grown on Matrigel (coating concentration 0.035 g/cm2 , BD #354234) in MEF-CM with the supplements described above.

    Techniques: Inhibition, Flow Cytometry, Cytometry

    Wnt/β-catenin activity is important for the self-renewal of naïve H1-4iLIF hESCs. Shown are graphs of the relative number of H1-4iLIF hESCs grown on Matrigel with or without (w/o) CHIR and with the indicated small molecules. Cells were

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Wnt/β-catenin signaling promotes self-renewal and inhibits the primed state transition in naïve human embryonic stem cells

    doi: 10.1073/pnas.1613849113

    Figure Lengend Snippet: Wnt/β-catenin activity is important for the self-renewal of naïve H1-4iLIF hESCs. Shown are graphs of the relative number of H1-4iLIF hESCs grown on Matrigel with or without (w/o) CHIR and with the indicated small molecules. Cells were

    Article Snippet: In some experiments, where indicated cells were grown on Matrigel (coating concentration 0.035 g/cm2 , BD #354234) in MEF-CM with the supplements described above.

    Techniques: Activity Assay

    Exogenous activation of Wnt/β-catenin signaling by recombinant Wnt3a partially restores self-renewal in naïve hESCs incubated with Wnt inhibitors. ( A and B ) Representative images of naïve ELF1 hESCs grown on Matrigel for 3 d (

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Wnt/β-catenin signaling promotes self-renewal and inhibits the primed state transition in naïve human embryonic stem cells

    doi: 10.1073/pnas.1613849113

    Figure Lengend Snippet: Exogenous activation of Wnt/β-catenin signaling by recombinant Wnt3a partially restores self-renewal in naïve hESCs incubated with Wnt inhibitors. ( A and B ) Representative images of naïve ELF1 hESCs grown on Matrigel for 3 d (

    Article Snippet: In some experiments, where indicated cells were grown on Matrigel (coating concentration 0.035 g/cm2 , BD #354234) in MEF-CM with the supplements described above.

    Techniques: Activation Assay, Recombinant, Incubation

    Inhibition of Wnt/β-catenin signaling reduces the gene expression of early differentiation markers. Shown are results of qRT-PCR–based analysis of gene expression in naïve ELF1 hESCs cultured on Matrigel with the indicated small

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Wnt/β-catenin signaling promotes self-renewal and inhibits the primed state transition in naïve human embryonic stem cells

    doi: 10.1073/pnas.1613849113

    Figure Lengend Snippet: Inhibition of Wnt/β-catenin signaling reduces the gene expression of early differentiation markers. Shown are results of qRT-PCR–based analysis of gene expression in naïve ELF1 hESCs cultured on Matrigel with the indicated small

    Article Snippet: In some experiments, where indicated cells were grown on Matrigel (coating concentration 0.035 g/cm2 , BD #354234) in MEF-CM with the supplements described above.

    Techniques: Inhibition, Expressing, Quantitative RT-PCR, Cell Culture

    Wnt inhibition leads to primed-like changes in naïve hESCs. ( A ) Results of qRT-PCR–based analysis of CRABP2 expression in ELF1-2iLIF and H1-4iLIF hESCs grown on Matrigel and treated with the indicated small molecules for 3 d. Fold changes

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Wnt/β-catenin signaling promotes self-renewal and inhibits the primed state transition in naïve human embryonic stem cells

    doi: 10.1073/pnas.1613849113

    Figure Lengend Snippet: Wnt inhibition leads to primed-like changes in naïve hESCs. ( A ) Results of qRT-PCR–based analysis of CRABP2 expression in ELF1-2iLIF and H1-4iLIF hESCs grown on Matrigel and treated with the indicated small molecules for 3 d. Fold changes

    Article Snippet: In some experiments, where indicated cells were grown on Matrigel (coating concentration 0.035 g/cm2 , BD #354234) in MEF-CM with the supplements described above.

    Techniques: Inhibition, Quantitative RT-PCR, Expressing

    Wnt/β-catenin activity is important for efficient self-renewal of naïve hESCs. ( A–C ) Graphs of the relative number of naïve ( A and B ) or primed ( C ) ELF1 hESCs grown on Matrigel with the indicated small molecules. Cells

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Wnt/β-catenin signaling promotes self-renewal and inhibits the primed state transition in naïve human embryonic stem cells

    doi: 10.1073/pnas.1613849113

    Figure Lengend Snippet: Wnt/β-catenin activity is important for efficient self-renewal of naïve hESCs. ( A–C ) Graphs of the relative number of naïve ( A and B ) or primed ( C ) ELF1 hESCs grown on Matrigel with the indicated small molecules. Cells

    Article Snippet: In some experiments, where indicated cells were grown on Matrigel (coating concentration 0.035 g/cm2 , BD #354234) in MEF-CM with the supplements described above.

    Techniques: Activity Assay

    Further examples for the strain stiffening effect. Single cells were seeded on Matrigel and the stiffness of the gel between the cells was measured with an AFM. As control, substrate stiffness in a cell-free area was also measured. The pseudo-coloured stiffness map shows the relative Young’s modulus normalized to the control area. From the overlay of the microscope Image and the stiffness map it can be seen that the cells formed a stiff bridge between them even before cellular protrusions form. In a time range from 30 min to 2 h, the cells used the stiffness line for locating each other and for contacting. Scale bar: 20 µm.

    Journal: bioRxiv

    Article Title: Cell based strain stiffening of a non-fibrous matrix as organizing principle for morphogenesis

    doi: 10.1101/816496

    Figure Lengend Snippet: Further examples for the strain stiffening effect. Single cells were seeded on Matrigel and the stiffness of the gel between the cells was measured with an AFM. As control, substrate stiffness in a cell-free area was also measured. The pseudo-coloured stiffness map shows the relative Young’s modulus normalized to the control area. From the overlay of the microscope Image and the stiffness map it can be seen that the cells formed a stiff bridge between them even before cellular protrusions form. In a time range from 30 min to 2 h, the cells used the stiffness line for locating each other and for contacting. Scale bar: 20 µm.

    Article Snippet: Matrigel (Corning, Amsterdam, Netherlands) was thawed on ice over night the day before use.

    Techniques: Microscopy

    Tube formation depends on active matrix deformation and substrate stiffness (A) Inhibition of cell contractility with 20 µM blebbistatin inhibited the formation of tubular structures. Scale bar: 100 µm (B) Upper left: Representative image of bead displacement by cells (red circles) on Matrigel as a dimensionless heatmap. Upper right: The analysis of the bead displacement over the first 3 h after seeding the cells on Matrigel showed that cells deform the matrix with a maximum around 40 µm. The energy (lower left) and radial strain (lower right) (C) HUVECs treatment with 20 µM blebbistatin reduced radial displacement, bulk deformation (20 x 10 4 cells/ml) were seeded on PAA or PDMS gels with different stiffness from 0.5 kPa to 5 kPa, coated with Matrigel, on Matrigel alone or on glass coated with Matrigel. At high (4 kPa and glass) stiffness values, the cells formed no network. On substrates with a stiffness between 0.5 and 1.5 kPa, HUVECs formed a tubular network like on the Matrigel control. Scale bar: 100 µm.

    Journal: bioRxiv

    Article Title: Cell based strain stiffening of a non-fibrous matrix as organizing principle for morphogenesis

    doi: 10.1101/816496

    Figure Lengend Snippet: Tube formation depends on active matrix deformation and substrate stiffness (A) Inhibition of cell contractility with 20 µM blebbistatin inhibited the formation of tubular structures. Scale bar: 100 µm (B) Upper left: Representative image of bead displacement by cells (red circles) on Matrigel as a dimensionless heatmap. Upper right: The analysis of the bead displacement over the first 3 h after seeding the cells on Matrigel showed that cells deform the matrix with a maximum around 40 µm. The energy (lower left) and radial strain (lower right) (C) HUVECs treatment with 20 µM blebbistatin reduced radial displacement, bulk deformation (20 x 10 4 cells/ml) were seeded on PAA or PDMS gels with different stiffness from 0.5 kPa to 5 kPa, coated with Matrigel, on Matrigel alone or on glass coated with Matrigel. At high (4 kPa and glass) stiffness values, the cells formed no network. On substrates with a stiffness between 0.5 and 1.5 kPa, HUVECs formed a tubular network like on the Matrigel control. Scale bar: 100 µm.

    Article Snippet: Matrigel (Corning, Amsterdam, Netherlands) was thawed on ice over night the day before use.

    Techniques: Inhibition

    Graphical Summary. While endothelial cells align to collagen fibres and move along them (left panels), they remodel the laminin based Matrigel (right panels): by contractile forces, strain stiffening occurs and the stiffened intercellular bridges are used as tracks for pattern formation.

    Journal: bioRxiv

    Article Title: Cell based strain stiffening of a non-fibrous matrix as organizing principle for morphogenesis

    doi: 10.1101/816496

    Figure Lengend Snippet: Graphical Summary. While endothelial cells align to collagen fibres and move along them (left panels), they remodel the laminin based Matrigel (right panels): by contractile forces, strain stiffening occurs and the stiffened intercellular bridges are used as tracks for pattern formation.

    Article Snippet: Matrigel (Corning, Amsterdam, Netherlands) was thawed on ice over night the day before use.

    Techniques:

    PDMS stiffness influences tube formation. (A) Schematic overview of the Matrigel printing on top of a PDMS gel. After activation of the PDMS with the plasma cleaner a thin layer with a thickness under 20 µm was printed on the PDMS. (B) PDMS stiffness in dependence of the rate of curing agent. (C) Immunostaining of HUVECs seeded on printed Matrigel on top of PDMS gels with different stiffness. If the stiffness was too high (4 kPa), cells did not form a tubular network, but simply spread on the surface. At 0.5 and 1.5 kPa cells interacted and tubes were formed. White arrows indicate the alignment of cells and fibrillary matrix. Scale bar: 100 µm.

    Journal: bioRxiv

    Article Title: Cell based strain stiffening of a non-fibrous matrix as organizing principle for morphogenesis

    doi: 10.1101/816496

    Figure Lengend Snippet: PDMS stiffness influences tube formation. (A) Schematic overview of the Matrigel printing on top of a PDMS gel. After activation of the PDMS with the plasma cleaner a thin layer with a thickness under 20 µm was printed on the PDMS. (B) PDMS stiffness in dependence of the rate of curing agent. (C) Immunostaining of HUVECs seeded on printed Matrigel on top of PDMS gels with different stiffness. If the stiffness was too high (4 kPa), cells did not form a tubular network, but simply spread on the surface. At 0.5 and 1.5 kPa cells interacted and tubes were formed. White arrows indicate the alignment of cells and fibrillary matrix. Scale bar: 100 µm.

    Article Snippet: Matrigel (Corning, Amsterdam, Netherlands) was thawed on ice over night the day before use.

    Techniques: Activation Assay, Immunostaining

    The initial finding phase depends on cell-cell distance. (A) HUVECs rapidly form cell-cell contacts when seeded on Matrigel. White arrows indicate the initial formation of the first cell-cell contacts in a time range from 1 to 2 h. Scale bar: 100 µm. (B) Tube formation after seeding different cell densities from 0.5 x 10 4 cells/ml to 20 x 10 4 cells/ml. At least 10 x 10 4 cells/ml were necessary to induce tube formation, while at lower cell numbers, HUVECs stayed separate or formed small group of cells. Images were taken after 6 h. Scale bar: 100 µm. (C) The cell densities inducing tube formation (blue curves) showed a higher mean squared displacement compared to the cell densities without tube formation (red curves). The normalized attraction and the velocity cross-correlation showed that cells can sense each other, to align and cluster, over a typical distance of at least 106 µm (vertical dotted line in the attraction graph; the typical interaction distance is extracted from an exponential fit with a constant offset, which is indicated in black). The typical time scale for the tube formation with 93 min is obtained by fitting the velocity autocorrelation with an exponential (indicated in black). A typical cell size is indicated by the grey region.

    Journal: bioRxiv

    Article Title: Cell based strain stiffening of a non-fibrous matrix as organizing principle for morphogenesis

    doi: 10.1101/816496

    Figure Lengend Snippet: The initial finding phase depends on cell-cell distance. (A) HUVECs rapidly form cell-cell contacts when seeded on Matrigel. White arrows indicate the initial formation of the first cell-cell contacts in a time range from 1 to 2 h. Scale bar: 100 µm. (B) Tube formation after seeding different cell densities from 0.5 x 10 4 cells/ml to 20 x 10 4 cells/ml. At least 10 x 10 4 cells/ml were necessary to induce tube formation, while at lower cell numbers, HUVECs stayed separate or formed small group of cells. Images were taken after 6 h. Scale bar: 100 µm. (C) The cell densities inducing tube formation (blue curves) showed a higher mean squared displacement compared to the cell densities without tube formation (red curves). The normalized attraction and the velocity cross-correlation showed that cells can sense each other, to align and cluster, over a typical distance of at least 106 µm (vertical dotted line in the attraction graph; the typical interaction distance is extracted from an exponential fit with a constant offset, which is indicated in black). The typical time scale for the tube formation with 93 min is obtained by fitting the velocity autocorrelation with an exponential (indicated in black). A typical cell size is indicated by the grey region.

    Article Snippet: Matrigel (Corning, Amsterdam, Netherlands) was thawed on ice over night the day before use.

    Techniques:

    Inhibition of proteolytic activity has no influence on tube formation. HUVECs were seeded on Matrigel and treated with 10 µM batimastat to inhibit a broad range of matrix metalloproteinases (MMPs). Images were taken after 6 h. Quantitative analysis of the number of tubes and nodes normalized to Control (Ctrl) showed that the activity of MMPs is not crucial for HUVEC tube formation. Two-tailed unpaired Student’s t test with Welch’s correction showed no significant differences. Scale bar: 100 µm.

    Journal: bioRxiv

    Article Title: Cell based strain stiffening of a non-fibrous matrix as organizing principle for morphogenesis

    doi: 10.1101/816496

    Figure Lengend Snippet: Inhibition of proteolytic activity has no influence on tube formation. HUVECs were seeded on Matrigel and treated with 10 µM batimastat to inhibit a broad range of matrix metalloproteinases (MMPs). Images were taken after 6 h. Quantitative analysis of the number of tubes and nodes normalized to Control (Ctrl) showed that the activity of MMPs is not crucial for HUVEC tube formation. Two-tailed unpaired Student’s t test with Welch’s correction showed no significant differences. Scale bar: 100 µm.

    Article Snippet: Matrigel (Corning, Amsterdam, Netherlands) was thawed on ice over night the day before use.

    Techniques: Inhibition, Activity Assay, Two Tailed Test

    Endothelial cells actively remodel ECM during early tube formation. Immunostaining of laminin and collagen IV in Matrigel without (first row, scale bar 10 µm), and with cells (second and third row, scale bar 50 µm). Cells time dependently re-arrange the matrix proteins so that fibres between cells are formed. This process depends on cell contractility and is inhibited by 20 µM blebbistatin (fourth and fifth row).

    Journal: bioRxiv

    Article Title: Cell based strain stiffening of a non-fibrous matrix as organizing principle for morphogenesis

    doi: 10.1101/816496

    Figure Lengend Snippet: Endothelial cells actively remodel ECM during early tube formation. Immunostaining of laminin and collagen IV in Matrigel without (first row, scale bar 10 µm), and with cells (second and third row, scale bar 50 µm). Cells time dependently re-arrange the matrix proteins so that fibres between cells are formed. This process depends on cell contractility and is inhibited by 20 µM blebbistatin (fourth and fifth row).

    Article Snippet: Matrigel (Corning, Amsterdam, Netherlands) was thawed on ice over night the day before use.

    Techniques: Immunostaining

    Cells change the local stiffness topography of the substrate. (A) Pseudo coloured AFM stiffness map of Matrigel and rat tail collagen I gel (Scale bar 5 µm): while the stiffness is evenly distributed in Matrigel, the collegen gel contains stiff fibres and soft interspaces. Right panel: the total reflection microscopy image of the collagen I gel shows the collagen fibre network (scale bar: 100 µm). Matrigel shows no fibrillary structure (data not shown). (B) Immunostaining of a collagen I gel in the presence of an endothelial cell. White arrows mark collagen fibres to which the cell is aligned (scale bar: 25 µm). Exemplary cell trajectories on Matrigel and collagen I over 20 h. Single cells on collagen migrated over a longer distance in all directions. The accumulated distance for 45 cells for each gel is normalized to the Matrigel and showed a significant increase on collagen I. Two-tailed unpaired Student’s t test with Welch’s correction, *P

    Journal: bioRxiv

    Article Title: Cell based strain stiffening of a non-fibrous matrix as organizing principle for morphogenesis

    doi: 10.1101/816496

    Figure Lengend Snippet: Cells change the local stiffness topography of the substrate. (A) Pseudo coloured AFM stiffness map of Matrigel and rat tail collagen I gel (Scale bar 5 µm): while the stiffness is evenly distributed in Matrigel, the collegen gel contains stiff fibres and soft interspaces. Right panel: the total reflection microscopy image of the collagen I gel shows the collagen fibre network (scale bar: 100 µm). Matrigel shows no fibrillary structure (data not shown). (B) Immunostaining of a collagen I gel in the presence of an endothelial cell. White arrows mark collagen fibres to which the cell is aligned (scale bar: 25 µm). Exemplary cell trajectories on Matrigel and collagen I over 20 h. Single cells on collagen migrated over a longer distance in all directions. The accumulated distance for 45 cells for each gel is normalized to the Matrigel and showed a significant increase on collagen I. Two-tailed unpaired Student’s t test with Welch’s correction, *P

    Article Snippet: Matrigel (Corning, Amsterdam, Netherlands) was thawed on ice over night the day before use.

    Techniques: Microscopy, Immunostaining, Two Tailed Test

    Laminin is a key factor for tube formation. (A) Upper row: immunostaining of laminin in Matrigel alone or after treatment with different rations of netrin-4 (Scale bar: 10 µm). Lower row: Tube formation on Matrigel and with different ratios of netrin-4 to laminin. Starting from a ratio of 1:1, netrin-4 changes the structure of the laminin network and influences tube formation. (B) Functionally blocking antibodies against integrins α1, α2, α3, α4, α5, α6 and αv were added (40 µg/ml). Blocking integrins, which preferentially bind to laminin (α1, α2, α3 and α6) inhibited tube formation, while blocking of mainly collagen and fibronectin binding integrins (α4, α5 and αv) did not affect endothelial tube formation. Scale bar: 100 µm.

    Journal: bioRxiv

    Article Title: Cell based strain stiffening of a non-fibrous matrix as organizing principle for morphogenesis

    doi: 10.1101/816496

    Figure Lengend Snippet: Laminin is a key factor for tube formation. (A) Upper row: immunostaining of laminin in Matrigel alone or after treatment with different rations of netrin-4 (Scale bar: 10 µm). Lower row: Tube formation on Matrigel and with different ratios of netrin-4 to laminin. Starting from a ratio of 1:1, netrin-4 changes the structure of the laminin network and influences tube formation. (B) Functionally blocking antibodies against integrins α1, α2, α3, α4, α5, α6 and αv were added (40 µg/ml). Blocking integrins, which preferentially bind to laminin (α1, α2, α3 and α6) inhibited tube formation, while blocking of mainly collagen and fibronectin binding integrins (α4, α5 and αv) did not affect endothelial tube formation. Scale bar: 100 µm.

    Article Snippet: Matrigel (Corning, Amsterdam, Netherlands) was thawed on ice over night the day before use.

    Techniques: Immunostaining, Blocking Assay, Binding Assay

    Role of the thickness of the Matrigel layer. (A) Matrigel was labelled with a fluorescent dye to measure the gel thickness by confocal microscopy. Different volumes of Matrigel from 1 to 10 µl were added to a PDMS gel, and the thickness was measured. To achieve a thickness of less than 20 µm, Matrigel was printed. Scale bar: 100 µm. (B) HUVECs were seeded on a stiff PDMS gel (70 kPa) coated with layers of Matrigel of a different thickness. On uncoated PDMS cells only adhered poorly and formed cell clusters. At a thickness under 20 µm, the cells sensed the stiffness of the underlying PDMS and formed no tubes. In all other cases, the cells did not sense the PDMS and formed the tubular network like on a thick Matrigel layer. Thus, printing of Matrigel was used for all further experiments. Scale bar: 100 µm.

    Journal: bioRxiv

    Article Title: Cell based strain stiffening of a non-fibrous matrix as organizing principle for morphogenesis

    doi: 10.1101/816496

    Figure Lengend Snippet: Role of the thickness of the Matrigel layer. (A) Matrigel was labelled with a fluorescent dye to measure the gel thickness by confocal microscopy. Different volumes of Matrigel from 1 to 10 µl were added to a PDMS gel, and the thickness was measured. To achieve a thickness of less than 20 µm, Matrigel was printed. Scale bar: 100 µm. (B) HUVECs were seeded on a stiff PDMS gel (70 kPa) coated with layers of Matrigel of a different thickness. On uncoated PDMS cells only adhered poorly and formed cell clusters. At a thickness under 20 µm, the cells sensed the stiffness of the underlying PDMS and formed no tubes. In all other cases, the cells did not sense the PDMS and formed the tubular network like on a thick Matrigel layer. Thus, printing of Matrigel was used for all further experiments. Scale bar: 100 µm.

    Article Snippet: Matrigel (Corning, Amsterdam, Netherlands) was thawed on ice over night the day before use.

    Techniques: Confocal Microscopy

    Tube formation on different hydrogel mixtures. (A) Young’s modulus of Matrigel, a rat tail collagen I gel (2 mg/ml), a mixture of laminin and collagen I in the ratio 6:1 (L:C 6:1) (4.8 mg/ml:0.8 mg/ml) and a laminin gel (6 mg/ml). The stiffness of Matrigel is significantly higher than the mixture L:C 6:1 or laminin, while there is no difference to collagen. One-way ANOVA, Dunnett’s test, compared to Matrigel, ***P

    Journal: bioRxiv

    Article Title: Cell based strain stiffening of a non-fibrous matrix as organizing principle for morphogenesis

    doi: 10.1101/816496

    Figure Lengend Snippet: Tube formation on different hydrogel mixtures. (A) Young’s modulus of Matrigel, a rat tail collagen I gel (2 mg/ml), a mixture of laminin and collagen I in the ratio 6:1 (L:C 6:1) (4.8 mg/ml:0.8 mg/ml) and a laminin gel (6 mg/ml). The stiffness of Matrigel is significantly higher than the mixture L:C 6:1 or laminin, while there is no difference to collagen. One-way ANOVA, Dunnett’s test, compared to Matrigel, ***P

    Article Snippet: Matrigel (Corning, Amsterdam, Netherlands) was thawed on ice over night the day before use.

    Techniques:

    LOXL2 expression induces ErbB2 activation through production of 2 O 2 . (A)Production of H 2 O 2 on Matrigel was measured as an activity read-out in the 10A cont and 10A L2 cells, by using a commercially available kit and following the manufacturer's instructions (Abcam). 10A L2 cells produced higher levels of H 2 O 2 , as expected, because of LOXL2 overexpression. (B) The 200 U/ml catalase (+cat) was added to remove H 2 O 2 produced by LOXL2. Western blotting revealed that pErbB2 was decreased in 10A L2 cells treated with catalase (L2+cat) in comparison with untreated cells (L2). (C) Addition of H 2 O 2 to 10A cont cells strongly induced ErbB2 activation. Taken together, these results indicate that, by removing H 2 O 2 , we can significantly abrogate ErbB2 activation in 10A L2 cells, and addition of H 2 O 2 activated ErbB2.

    Journal: Breast Cancer Research : BCR

    Article Title: LOXL2 induces aberrant acinar morphogenesis via ErbB2 signaling

    doi: 10.1186/bcr3461

    Figure Lengend Snippet: LOXL2 expression induces ErbB2 activation through production of 2 O 2 . (A)Production of H 2 O 2 on Matrigel was measured as an activity read-out in the 10A cont and 10A L2 cells, by using a commercially available kit and following the manufacturer's instructions (Abcam). 10A L2 cells produced higher levels of H 2 O 2 , as expected, because of LOXL2 overexpression. (B) The 200 U/ml catalase (+cat) was added to remove H 2 O 2 produced by LOXL2. Western blotting revealed that pErbB2 was decreased in 10A L2 cells treated with catalase (L2+cat) in comparison with untreated cells (L2). (C) Addition of H 2 O 2 to 10A cont cells strongly induced ErbB2 activation. Taken together, these results indicate that, by removing H 2 O 2 , we can significantly abrogate ErbB2 activation in 10A L2 cells, and addition of H 2 O 2 activated ErbB2.

    Article Snippet: Acini assay 10A cont and 10A L2 cells were plated on top of a thin layer of 50-μl Matrigel in eight-well chamber slides, at a density of 10,000 cells/ml in 400 μl of 2% Growth Factor Reduced Matrigel without phenol red (BD Biosciences) in DMEM/F-12 media supplemented with 2% HS, 0.5 μg/ml hydrocortisone, 10 μg/ml insulin, 100 ng/ml cholera toxin, and 20 ng/ml EGF (Gibco/Invitrogen).

    Techniques: Expressing, Activation Assay, Activity Assay, Produced, Over Expression, Western Blot

    LOXL2 promotes invasion of normal breast epithelial cells; this effect is abrogated with ErbB2 inhibition . (A) The 10A L2 cells exhibited increased invasion through Matrigel compared with 10A cont cells in Transwell invasion assays. (B) Herceptin treatment reduced the invasive ability of the 10A L2 cells to a level comparable to 10A cont cells. Human IgG treatment had no effect on the invasiveness of the manipulated 10A cells. (C) Lapatinib treatment of the 10A L2 cells greatly reduced the invasiveness of the cells to a level comparable to the 10A cont cells. DMSO treatment had no effect on the invasive properties of the manipulated 10A cells.

    Journal: Breast Cancer Research : BCR

    Article Title: LOXL2 induces aberrant acinar morphogenesis via ErbB2 signaling

    doi: 10.1186/bcr3461

    Figure Lengend Snippet: LOXL2 promotes invasion of normal breast epithelial cells; this effect is abrogated with ErbB2 inhibition . (A) The 10A L2 cells exhibited increased invasion through Matrigel compared with 10A cont cells in Transwell invasion assays. (B) Herceptin treatment reduced the invasive ability of the 10A L2 cells to a level comparable to 10A cont cells. Human IgG treatment had no effect on the invasiveness of the manipulated 10A cells. (C) Lapatinib treatment of the 10A L2 cells greatly reduced the invasiveness of the cells to a level comparable to the 10A cont cells. DMSO treatment had no effect on the invasive properties of the manipulated 10A cells.

    Article Snippet: Acini assay 10A cont and 10A L2 cells were plated on top of a thin layer of 50-μl Matrigel in eight-well chamber slides, at a density of 10,000 cells/ml in 400 μl of 2% Growth Factor Reduced Matrigel without phenol red (BD Biosciences) in DMEM/F-12 media supplemented with 2% HS, 0.5 μg/ml hydrocortisone, 10 μg/ml insulin, 100 ng/ml cholera toxin, and 20 ng/ml EGF (Gibco/Invitrogen).

    Techniques: Inhibition

    LOXL2 expression disrupts normal breast epithelial acini formation . 10A cont and 10A L2 cells were plated on top of a thin layer of 50 μl Matrigel in eight-well chamber slides in a suspension of Matrigel/culture media mix to investigate the acinar morphogenesis of these cells. The cultures were allowed to grow for 8, 10, and 13 days, and then fixed and stained with primary antibodies, as described in the different panels. All data were based on at least three independent experimental repeats. Scale bar, 20 μm. (A) Immunofluorescence staining of acini with anti-activated caspase-3 antibody to detect apoptotic cells on day 8 reveals decreased apoptosis in the central cells of the L2 acini. Representative images of activated caspase-3 staining in acini for each cell line are shown. Quantification represented average percentage ± standard error of acini containing activated caspase-3-positive cells ( P = 0.0007). (B) Immunofluorescence staining of acini with anti-Ki67 antibody to detect proliferating cells on day 13 reveals that L2 acini had more proliferative cells. Representative images of Ki67-positive acini from each cell line are shown. Quantification represented average percentage ± standard error of acini containing Ki67-positive cells. ( P = 0.0007). (C) Immunofluorescence staining of acini with anti-GM130 antibody to assess cell polarity on day 10. Representative images of GM130 staining in acini for each cell line are shown. Quantification represented average percentage ± standard error of acini forming a regular ring structure, as assessed by GM130 staining ( P = 0.0005; n = 80 acini for each cell line per repeat). (D) Quantification of average percentage of acini at day 13 with evacuated lumens ± standard error. Acini with evacuated lumens were defined as having no more than 20% of total number of cells, as well as Ki67-positive cells present in the center ( P = 0.013).

    Journal: Breast Cancer Research : BCR

    Article Title: LOXL2 induces aberrant acinar morphogenesis via ErbB2 signaling

    doi: 10.1186/bcr3461

    Figure Lengend Snippet: LOXL2 expression disrupts normal breast epithelial acini formation . 10A cont and 10A L2 cells were plated on top of a thin layer of 50 μl Matrigel in eight-well chamber slides in a suspension of Matrigel/culture media mix to investigate the acinar morphogenesis of these cells. The cultures were allowed to grow for 8, 10, and 13 days, and then fixed and stained with primary antibodies, as described in the different panels. All data were based on at least three independent experimental repeats. Scale bar, 20 μm. (A) Immunofluorescence staining of acini with anti-activated caspase-3 antibody to detect apoptotic cells on day 8 reveals decreased apoptosis in the central cells of the L2 acini. Representative images of activated caspase-3 staining in acini for each cell line are shown. Quantification represented average percentage ± standard error of acini containing activated caspase-3-positive cells ( P = 0.0007). (B) Immunofluorescence staining of acini with anti-Ki67 antibody to detect proliferating cells on day 13 reveals that L2 acini had more proliferative cells. Representative images of Ki67-positive acini from each cell line are shown. Quantification represented average percentage ± standard error of acini containing Ki67-positive cells. ( P = 0.0007). (C) Immunofluorescence staining of acini with anti-GM130 antibody to assess cell polarity on day 10. Representative images of GM130 staining in acini for each cell line are shown. Quantification represented average percentage ± standard error of acini forming a regular ring structure, as assessed by GM130 staining ( P = 0.0005; n = 80 acini for each cell line per repeat). (D) Quantification of average percentage of acini at day 13 with evacuated lumens ± standard error. Acini with evacuated lumens were defined as having no more than 20% of total number of cells, as well as Ki67-positive cells present in the center ( P = 0.013).

    Article Snippet: Acini assay 10A cont and 10A L2 cells were plated on top of a thin layer of 50-μl Matrigel in eight-well chamber slides, at a density of 10,000 cells/ml in 400 μl of 2% Growth Factor Reduced Matrigel without phenol red (BD Biosciences) in DMEM/F-12 media supplemented with 2% HS, 0.5 μg/ml hydrocortisone, 10 μg/ml insulin, 100 ng/ml cholera toxin, and 20 ng/ml EGF (Gibco/Invitrogen).

    Techniques: Expressing, Staining, Immunofluorescence

    LOXL2 expression in MCF10A cells increases phosphorylation of ErbB2 . Cells were plated on a thin layer of Matrigel and serum-starved for 3 hours before being subjected to serum-blasting and subsequent lysis of cells. (A) Western blotting revealed that phospho-ErbB2 was elevated in 10A L2 cells when compared with 10A cont cells, whereas total ErbB2 levels were equivalent in the two lines, suggesting increased phosphorylation of ErbB2 in the 10A L2 cells. Densitometry analysis was calculated for pErbB2 levels relative to total ErbB2 and revealed a significant increase in 10A L2 cells ( P = 0.0241). The levels of phospho-Akt and phospho-Erk1/2 were also elevated in 10A L2 cells. (B) The 10A cont cells were subjected to 16-hour treatment with 50 nM recombinant human LOXL2 (rhLOXL2; R D Systems) followed by serum-starvation and serum-blasting (10A cont + rhLOXL2). Western-blotting analysis showed that in 10A cont treated with rhLOXL2, phospho-ErbB2 level was increased to a greater extent than 10A L2 cells when compared with sham-treated 10A cont cells. Densitometry analysis was calculated for pErbB2 levels relative to total ErbB2 and revealed significant increase in 10A cont + rhLOXL2 cells ( P = 0.0438). This suggested that extracellular recombinant LOXL2 was capable of activating the ErbB2 receptor in 10A cells.

    Journal: Breast Cancer Research : BCR

    Article Title: LOXL2 induces aberrant acinar morphogenesis via ErbB2 signaling

    doi: 10.1186/bcr3461

    Figure Lengend Snippet: LOXL2 expression in MCF10A cells increases phosphorylation of ErbB2 . Cells were plated on a thin layer of Matrigel and serum-starved for 3 hours before being subjected to serum-blasting and subsequent lysis of cells. (A) Western blotting revealed that phospho-ErbB2 was elevated in 10A L2 cells when compared with 10A cont cells, whereas total ErbB2 levels were equivalent in the two lines, suggesting increased phosphorylation of ErbB2 in the 10A L2 cells. Densitometry analysis was calculated for pErbB2 levels relative to total ErbB2 and revealed a significant increase in 10A L2 cells ( P = 0.0241). The levels of phospho-Akt and phospho-Erk1/2 were also elevated in 10A L2 cells. (B) The 10A cont cells were subjected to 16-hour treatment with 50 nM recombinant human LOXL2 (rhLOXL2; R D Systems) followed by serum-starvation and serum-blasting (10A cont + rhLOXL2). Western-blotting analysis showed that in 10A cont treated with rhLOXL2, phospho-ErbB2 level was increased to a greater extent than 10A L2 cells when compared with sham-treated 10A cont cells. Densitometry analysis was calculated for pErbB2 levels relative to total ErbB2 and revealed significant increase in 10A cont + rhLOXL2 cells ( P = 0.0438). This suggested that extracellular recombinant LOXL2 was capable of activating the ErbB2 receptor in 10A cells.

    Article Snippet: Acini assay 10A cont and 10A L2 cells were plated on top of a thin layer of 50-μl Matrigel in eight-well chamber slides, at a density of 10,000 cells/ml in 400 μl of 2% Growth Factor Reduced Matrigel without phenol red (BD Biosciences) in DMEM/F-12 media supplemented with 2% HS, 0.5 μg/ml hydrocortisone, 10 μg/ml insulin, 100 ng/ml cholera toxin, and 20 ng/ml EGF (Gibco/Invitrogen).

    Techniques: Expressing, Lysis, Western Blot, Recombinant

    LOXL2-expressing MCF10A cells form ErbB2-dependent branching structures on matrix . (A) When plated on Matrigel-coated plates, 10A cont cells remained small and discrete, whereas 10A L2 cells formed extensive branching structures. Quantification of branch-points revealed that 10A L2 cells have significantly more branch-points (right panel; P = 0.00004). (B) Herceptin treatment of the 10A L2 cells significantly decreased the degree of branching of these cells to that seen for controls (right panel; P = 0.0167 and P = 0.0398, respectively). (C) Lapatinib treatment of the 10A L2 cells significantly abrogated the branching ability of these cells to that seen for controls (right panel; P = 0.0007 and P = 0.0003, respectively).

    Journal: Breast Cancer Research : BCR

    Article Title: LOXL2 induces aberrant acinar morphogenesis via ErbB2 signaling

    doi: 10.1186/bcr3461

    Figure Lengend Snippet: LOXL2-expressing MCF10A cells form ErbB2-dependent branching structures on matrix . (A) When plated on Matrigel-coated plates, 10A cont cells remained small and discrete, whereas 10A L2 cells formed extensive branching structures. Quantification of branch-points revealed that 10A L2 cells have significantly more branch-points (right panel; P = 0.00004). (B) Herceptin treatment of the 10A L2 cells significantly decreased the degree of branching of these cells to that seen for controls (right panel; P = 0.0167 and P = 0.0398, respectively). (C) Lapatinib treatment of the 10A L2 cells significantly abrogated the branching ability of these cells to that seen for controls (right panel; P = 0.0007 and P = 0.0003, respectively).

    Article Snippet: Acini assay 10A cont and 10A L2 cells were plated on top of a thin layer of 50-μl Matrigel in eight-well chamber slides, at a density of 10,000 cells/ml in 400 μl of 2% Growth Factor Reduced Matrigel without phenol red (BD Biosciences) in DMEM/F-12 media supplemented with 2% HS, 0.5 μg/ml hydrocortisone, 10 μg/ml insulin, 100 ng/ml cholera toxin, and 20 ng/ml EGF (Gibco/Invitrogen).

    Techniques: Expressing

    Manipulation of LOXL2 expression levels in human MCF10A normal mammary epithelial cells . (A) Western blot of secreted LOXL2 in CM generated from MDA MB-231 cells (MDA 231), MCF10A cells infected with LOXL2 (10A L2) and vector alone (10A cont) revealed that LOXL2 protein expression was upregulated in the 10A L2 cells to a level similar to that detected in MDA cells. β-Actin was used as a loading control. (B) Quantitative real-time PCR (qRT-PCR) of LOXL2 mRNA levels in manipulated MCF10A cells showed that LOXL2 mRNA levels were upregulated in 10A L2 cells. P = 0.009. (C) Morphologies of the manipulated MCF10A cells compared with WT cells showed that upregulation of LOXL2 did not produce significant alterations on cells plated on 2D tissue-culture plastic. Cells were viewed under a microscope (Leica DM1L), and representative images were taken. (D) 2D MTS proliferation assay of manipulated MCF10A cells suggested that when cultured on plastic, LOXL2 expression did not alter proliferation of the 10A cells. Error bars represent SEM for three independent experiments. (E) 3D MTS proliferation assay of manipulated MCF10A cells with manipulated LOXL2 expression plated within Matrigel suspension suggested that increased LOXL2 expression increases proliferation of the 10A cells in 3D. Error bars represent SEM for three independent experiments. P = 0.034 for day 6.

    Journal: Breast Cancer Research : BCR

    Article Title: LOXL2 induces aberrant acinar morphogenesis via ErbB2 signaling

    doi: 10.1186/bcr3461

    Figure Lengend Snippet: Manipulation of LOXL2 expression levels in human MCF10A normal mammary epithelial cells . (A) Western blot of secreted LOXL2 in CM generated from MDA MB-231 cells (MDA 231), MCF10A cells infected with LOXL2 (10A L2) and vector alone (10A cont) revealed that LOXL2 protein expression was upregulated in the 10A L2 cells to a level similar to that detected in MDA cells. β-Actin was used as a loading control. (B) Quantitative real-time PCR (qRT-PCR) of LOXL2 mRNA levels in manipulated MCF10A cells showed that LOXL2 mRNA levels were upregulated in 10A L2 cells. P = 0.009. (C) Morphologies of the manipulated MCF10A cells compared with WT cells showed that upregulation of LOXL2 did not produce significant alterations on cells plated on 2D tissue-culture plastic. Cells were viewed under a microscope (Leica DM1L), and representative images were taken. (D) 2D MTS proliferation assay of manipulated MCF10A cells suggested that when cultured on plastic, LOXL2 expression did not alter proliferation of the 10A cells. Error bars represent SEM for three independent experiments. (E) 3D MTS proliferation assay of manipulated MCF10A cells with manipulated LOXL2 expression plated within Matrigel suspension suggested that increased LOXL2 expression increases proliferation of the 10A cells in 3D. Error bars represent SEM for three independent experiments. P = 0.034 for day 6.

    Article Snippet: Acini assay 10A cont and 10A L2 cells were plated on top of a thin layer of 50-μl Matrigel in eight-well chamber slides, at a density of 10,000 cells/ml in 400 μl of 2% Growth Factor Reduced Matrigel without phenol red (BD Biosciences) in DMEM/F-12 media supplemented with 2% HS, 0.5 μg/ml hydrocortisone, 10 μg/ml insulin, 100 ng/ml cholera toxin, and 20 ng/ml EGF (Gibco/Invitrogen).

    Techniques: Expressing, Western Blot, Generated, Multiple Displacement Amplification, Infection, Plasmid Preparation, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Microscopy, Proliferation Assay, Cell Culture

    The E-cadherin expression of EpH4 cells 7 days after incubation of EpH4 cells with MM, GM, and matrigel-coated GM or without microspheres and EpH4 cells were cultured by the conventional 3D and 2D monolayer methods in the absence (□) or presence of PI3K inhibitor LY294002 (■). The number of EpH4 cells added initially was 1 × 10 4 /well while that of microspheres was 1 × 10 2 or 1 × 10 3 /well. *, p

    Journal: Regenerative Therapy

    Article Title: Preparation of epithelial cell aggregates incorporating matrigel microspheres to enhance proliferation and differentiation of epithelial cells

    doi: 10.1016/j.reth.2017.07.001

    Figure Lengend Snippet: The E-cadherin expression of EpH4 cells 7 days after incubation of EpH4 cells with MM, GM, and matrigel-coated GM or without microspheres and EpH4 cells were cultured by the conventional 3D and 2D monolayer methods in the absence (□) or presence of PI3K inhibitor LY294002 (■). The number of EpH4 cells added initially was 1 × 10 4 /well while that of microspheres was 1 × 10 2 or 1 × 10 3 /well. *, p

    Article Snippet: Briefly, 1.0 ml of 10 vol% aqueous Becton, Dickinson and Company (BD) Matrigel™ Basement Membrane Matrix (BD Biosciences, Inc., Franklin Lakes, America) solution was prepared at 4 °C.

    Techniques: Expressing, Incubation, Cell Culture

    The β-casein expression of EpH4 cells 4 (□), 7 ( Image 1 ), and 14 days (■) after incubation of EpH4 cells with MM, GM, and matrigel-coated GM or without microspheres. EpH4 cells were cultured by the conventional 3D and 2D monolayer methods. The number of EpH4 cells added initially was 1 × 10 4 /well while that of microspheres was 1 × 10 2 or 1 × 10 3 /well. *, p

    Journal: Regenerative Therapy

    Article Title: Preparation of epithelial cell aggregates incorporating matrigel microspheres to enhance proliferation and differentiation of epithelial cells

    doi: 10.1016/j.reth.2017.07.001

    Figure Lengend Snippet: The β-casein expression of EpH4 cells 4 (□), 7 ( Image 1 ), and 14 days (■) after incubation of EpH4 cells with MM, GM, and matrigel-coated GM or without microspheres. EpH4 cells were cultured by the conventional 3D and 2D monolayer methods. The number of EpH4 cells added initially was 1 × 10 4 /well while that of microspheres was 1 × 10 2 or 1 × 10 3 /well. *, p

    Article Snippet: Briefly, 1.0 ml of 10 vol% aqueous Becton, Dickinson and Company (BD) Matrigel™ Basement Membrane Matrix (BD Biosciences, Inc., Franklin Lakes, America) solution was prepared at 4 °C.

    Techniques: Expressing, Incubation, Cell Culture

    The number of live EpH4 cells 7 days after incubation of EpH4 cells with MM, GM, and matrigel-coated GM or without microspheres. EpH4 cells were cultured by the conventional 3D and 2D monolayer methods. The number of EpH4 cells added initially was 1 × 10 4 /well while that of microspheres was 1 × 10 2 or 1 × 10 3 /well. *, p

    Journal: Regenerative Therapy

    Article Title: Preparation of epithelial cell aggregates incorporating matrigel microspheres to enhance proliferation and differentiation of epithelial cells

    doi: 10.1016/j.reth.2017.07.001

    Figure Lengend Snippet: The number of live EpH4 cells 7 days after incubation of EpH4 cells with MM, GM, and matrigel-coated GM or without microspheres. EpH4 cells were cultured by the conventional 3D and 2D monolayer methods. The number of EpH4 cells added initially was 1 × 10 4 /well while that of microspheres was 1 × 10 2 or 1 × 10 3 /well. *, p

    Article Snippet: Briefly, 1.0 ml of 10 vol% aqueous Becton, Dickinson and Company (BD) Matrigel™ Basement Membrane Matrix (BD Biosciences, Inc., Franklin Lakes, America) solution was prepared at 4 °C.

    Techniques: Incubation, Cell Culture

    Confocal microscopic pictures of β-casein expression of EpH4 cells 7 days after incubation of EpH4 cells with MM, GM, and matrigel-coated GM or without microspheres. EpH4 cells were cultured by the conventional 3D and 2D monolayer methods. The number of EpH4 cells added initially was 1 × 10 4 /well while that of microspheres was 1 × 10 2 or 1 × 10 3 /well. Scale bar. 100 μm.

    Journal: Regenerative Therapy

    Article Title: Preparation of epithelial cell aggregates incorporating matrigel microspheres to enhance proliferation and differentiation of epithelial cells

    doi: 10.1016/j.reth.2017.07.001

    Figure Lengend Snippet: Confocal microscopic pictures of β-casein expression of EpH4 cells 7 days after incubation of EpH4 cells with MM, GM, and matrigel-coated GM or without microspheres. EpH4 cells were cultured by the conventional 3D and 2D monolayer methods. The number of EpH4 cells added initially was 1 × 10 4 /well while that of microspheres was 1 × 10 2 or 1 × 10 3 /well. Scale bar. 100 μm.

    Article Snippet: Briefly, 1.0 ml of 10 vol% aqueous Becton, Dickinson and Company (BD) Matrigel™ Basement Membrane Matrix (BD Biosciences, Inc., Franklin Lakes, America) solution was prepared at 4 °C.

    Techniques: Expressing, Incubation, Cell Culture

    The l -lactic acid/glucose ratio of live EpH4 cells 7 days after incubation of EpH4 cells with MM, GM, and matrigel-coated GM or without microspheres. EpH4 cells were cultured by the conventional 3D and 2D monolayer methods. The number of EpH4 cells added initially was 1 × 10 4 /well while that of microspheres was 1 × 10 2 or 1 × 10 3 /well.

    Journal: Regenerative Therapy

    Article Title: Preparation of epithelial cell aggregates incorporating matrigel microspheres to enhance proliferation and differentiation of epithelial cells

    doi: 10.1016/j.reth.2017.07.001

    Figure Lengend Snippet: The l -lactic acid/glucose ratio of live EpH4 cells 7 days after incubation of EpH4 cells with MM, GM, and matrigel-coated GM or without microspheres. EpH4 cells were cultured by the conventional 3D and 2D monolayer methods. The number of EpH4 cells added initially was 1 × 10 4 /well while that of microspheres was 1 × 10 2 or 1 × 10 3 /well.

    Article Snippet: Briefly, 1.0 ml of 10 vol% aqueous Becton, Dickinson and Company (BD) Matrigel™ Basement Membrane Matrix (BD Biosciences, Inc., Franklin Lakes, America) solution was prepared at 4 °C.

    Techniques: Incubation, Cell Culture

    Confocal microscopic pictures of laminin expression of EpH4 cells 7 days after incubation of EpH4 cells with MM, GM, and matrigel-coated GM or without microspheres. EpH4 cells were cultured by the conventional 3D and 2D monolayer methods. The number of EpH4 cells added initially was 1 × 10 4 /well while that of microspheres was 1 × 10 2 or 1 × 10 3 /well. Scale bar. 100 μm.

    Journal: Regenerative Therapy

    Article Title: Preparation of epithelial cell aggregates incorporating matrigel microspheres to enhance proliferation and differentiation of epithelial cells

    doi: 10.1016/j.reth.2017.07.001

    Figure Lengend Snippet: Confocal microscopic pictures of laminin expression of EpH4 cells 7 days after incubation of EpH4 cells with MM, GM, and matrigel-coated GM or without microspheres. EpH4 cells were cultured by the conventional 3D and 2D monolayer methods. The number of EpH4 cells added initially was 1 × 10 4 /well while that of microspheres was 1 × 10 2 or 1 × 10 3 /well. Scale bar. 100 μm.

    Article Snippet: Briefly, 1.0 ml of 10 vol% aqueous Becton, Dickinson and Company (BD) Matrigel™ Basement Membrane Matrix (BD Biosciences, Inc., Franklin Lakes, America) solution was prepared at 4 °C.

    Techniques: Expressing, Incubation, Cell Culture

    A confocal microscopic picture of matrigel-coated GM dispersed in water. The GM were dehydrothermally crosslinked for 18 h at 140 °C. The laminin of matrigel was immunostained. In green. Scale bar. 100 μm.

    Journal: Regenerative Therapy

    Article Title: Preparation of epithelial cell aggregates incorporating matrigel microspheres to enhance proliferation and differentiation of epithelial cells

    doi: 10.1016/j.reth.2017.07.001

    Figure Lengend Snippet: A confocal microscopic picture of matrigel-coated GM dispersed in water. The GM were dehydrothermally crosslinked for 18 h at 140 °C. The laminin of matrigel was immunostained. In green. Scale bar. 100 μm.

    Article Snippet: Briefly, 1.0 ml of 10 vol% aqueous Becton, Dickinson and Company (BD) Matrigel™ Basement Membrane Matrix (BD Biosciences, Inc., Franklin Lakes, America) solution was prepared at 4 °C.

    Techniques:

    The β-casein expression of EpH4 cells 7 days after incubation of EpH4 cells with MM, GM, and matrigel-coated GM or without microspheres and EpH4 cells were cultured by the conventional 3D and 2D monolayer methods in the absence (□) or presence of PI3K inhibitor LY294002 (■). The number of EpH4 cells added initially was 1 × 10 4 /well while that of microspheres was 1 × 10 2 or 1 × 10 3 /well. *, p

    Journal: Regenerative Therapy

    Article Title: Preparation of epithelial cell aggregates incorporating matrigel microspheres to enhance proliferation and differentiation of epithelial cells

    doi: 10.1016/j.reth.2017.07.001

    Figure Lengend Snippet: The β-casein expression of EpH4 cells 7 days after incubation of EpH4 cells with MM, GM, and matrigel-coated GM or without microspheres and EpH4 cells were cultured by the conventional 3D and 2D monolayer methods in the absence (□) or presence of PI3K inhibitor LY294002 (■). The number of EpH4 cells added initially was 1 × 10 4 /well while that of microspheres was 1 × 10 2 or 1 × 10 3 /well. *, p

    Article Snippet: Briefly, 1.0 ml of 10 vol% aqueous Becton, Dickinson and Company (BD) Matrigel™ Basement Membrane Matrix (BD Biosciences, Inc., Franklin Lakes, America) solution was prepared at 4 °C.

    Techniques: Expressing, Incubation, Cell Culture

    Live/dead assay of EpH4 cell aggregates 7 days after incubation of EpH4 cells with MM, GM, and matrigel-coated GM or without microspheres. EpH4 cells were cultured by the conventional 3D and 2D monolayer methods. The number of EpH4 cells added initially was 1 × 10 4 /well while that of microspheres was 1 × 10 2 or 1 × 10 3 /well. Scale bar. 100 μm.

    Journal: Regenerative Therapy

    Article Title: Preparation of epithelial cell aggregates incorporating matrigel microspheres to enhance proliferation and differentiation of epithelial cells

    doi: 10.1016/j.reth.2017.07.001

    Figure Lengend Snippet: Live/dead assay of EpH4 cell aggregates 7 days after incubation of EpH4 cells with MM, GM, and matrigel-coated GM or without microspheres. EpH4 cells were cultured by the conventional 3D and 2D monolayer methods. The number of EpH4 cells added initially was 1 × 10 4 /well while that of microspheres was 1 × 10 2 or 1 × 10 3 /well. Scale bar. 100 μm.

    Article Snippet: Briefly, 1.0 ml of 10 vol% aqueous Becton, Dickinson and Company (BD) Matrigel™ Basement Membrane Matrix (BD Biosciences, Inc., Franklin Lakes, America) solution was prepared at 4 °C.

    Techniques: Live Dead Assay, Incubation, Cell Culture

    Light microscopic pictures of EpH4 cell aggregates 7 days after incubation of EpH4 cells with MM, GM, and matrigel-coated GM or without microspheres. EpH4 cells were cultured as by the conventional 3D and 2D monolayer culture methods. The number of EpH4 cells added initially was 1 × 10 4 /well while that of microspheres was 1 × 10 2 , 1 × 10 3 or 1 × 10 4 /well. Scale bar. 100 μm.

    Journal: Regenerative Therapy

    Article Title: Preparation of epithelial cell aggregates incorporating matrigel microspheres to enhance proliferation and differentiation of epithelial cells

    doi: 10.1016/j.reth.2017.07.001

    Figure Lengend Snippet: Light microscopic pictures of EpH4 cell aggregates 7 days after incubation of EpH4 cells with MM, GM, and matrigel-coated GM or without microspheres. EpH4 cells were cultured as by the conventional 3D and 2D monolayer culture methods. The number of EpH4 cells added initially was 1 × 10 4 /well while that of microspheres was 1 × 10 2 , 1 × 10 3 or 1 × 10 4 /well. Scale bar. 100 μm.

    Article Snippet: Briefly, 1.0 ml of 10 vol% aqueous Becton, Dickinson and Company (BD) Matrigel™ Basement Membrane Matrix (BD Biosciences, Inc., Franklin Lakes, America) solution was prepared at 4 °C.

    Techniques: Incubation, Cell Culture