Journal: ACS Applied Materials & Interfaces

Article Title: Systematic Comparison of Commercial Hydrogels Revealed That a Synergy of Laminin and Strain-Stiffening Promotes Directed Migration of Neural Cells

doi: 10.1021/acsami.2c20040

Figure Lengend Snippet: DRG neuron cultures seeded in hydrogels and CTRL. (a) Phase contrast micrographs of DRG neuron cultures on (1) PDL/laminin-coated cell culture dishes (CTRL) and in hydrogels (2) PuraMatrix, (3) Cultrex BME, and (4) Novatach. (b) Representative confocal micrographs of cultures stained for TUJ1 in green, S100 in red, vimentin (VIME) in orange, and DAPI in blue. Images in the left corner are representative micrographs of DAPI + cell nuclei of a single DRG neuron, indicated by white arrows. Enlargements of the neurite networks show TUJ1 and S100 stainings to assess neurite/SC co-localization indicated by white arrowheads. (c) Diagram depicts the mean ± SD percentage of S100 (SC) and TUJ1 (DRG neurons) overlap ( n = 3). (d) Diagram depicts the mean ± SD nuclei size of DRG neurons in μm 2 for the four conditions ( n = 3). (e) Diagram depicts the mean ± SD nuclei roundness ( n = 3). (f) Diagram depicts the mean ± SD number of primary neurites ( n = 3). (g) Diagram depicts the mean ± SD number of BPs per 500 μm neurite length ( n = 3). (h) Diagram depicts the mean ± SD neurite thickness in μm ( n = 3). * p -value < 0.05, ** p -value < 0.01, *** p -value < 0.001.

Article Snippet: The synthetic, polypeptide hydrogel PuraMatrix (Corning), the basement membrane extract hydrogel Cultrex 3D Cell Culture Matrix Basement Membrane Extract Reduced Growth Factor (Trevigen), the laminin hydrogel Cultrex 3D Cell Culture Matrix Laminin I, the collagen hydrogel Cultrex 3D Cell Culture Matrix Rat Tail Collagen I, and the alginate peptide-coupled hydrogel Novatach (Novamatrix) were diluted according to the manufacturer’s recommendations.

Techniques: Cell Culture, Staining

Journal: ACS Applied Materials & Interfaces

Article Title: Systematic Comparison of Commercial Hydrogels Revealed That a Synergy of Laminin and Strain-Stiffening Promotes Directed Migration of Neural Cells

doi: 10.1021/acsami.2c20040

Figure Lengend Snippet: Evaluation of SC and FB morphology, proliferation, and viability. (a) Representative phase contrast micrographs of SC cultures on (a1) poly- l -lysine (PLL)/laminin-coated cell culture dishes (CTRL), (a2) PuraMatrix, (a3) Cultrex BME, and (a4) Novatach. Hydrogel residues of PuraMatrix were free-floating in the medium [arrowheads in (a2)]. (b) Representative confocal micrographs of SCs stained for S100 in green, VIME in gray, EdU in red, and DAPI in blue. The arrowheads indicate proliferating SCs. Small images in the right corner are representative micrographs of a DAPI + cell nucleus of a single SC, indicated by white arrows. (c) Diagram depicts the mean ± SD length/width ratio of SCs on PLL/laminin (CTRL) and hydrogels ( n = 7). (d) Diagram depicts the mean ± SD nuclei size of SCs in μm 2 for the four groups ( n = 7). (e) Diagram depicts the roundness of SC nuclei on PLL/laminin and hydrogels ± SD ( n = 7). (f) Diagram shows the mean ± SD percentage of S100 + cells (SCs) for each group ( n = 6). (g) Diagram depicts the mean ± SD percentage of S100 + /EdU + cells (proliferating SCs) in each condition ( n = 7). (h) Diagram depicts the percentage of SCs negative for propidium iodide (PI) in each condition ± SD ( n = 6). (i) Representative phase contrast micrographs of FB cultures on (i1) uncoated cell culture wells (CTRL), (i2) PuraMatrix, (i3) Cultrex BME, and (i4) Novatach. (j) Representative confocal micrographs of FBs stained for THY1 in cyan, NGFR in pink, EdU in red, and DAPI in gray. The arrowhead indicates proliferating FBs, and the pink arrows indicate NGFR + SCs. Small images in the right corner of the image are representative micrographs of a DAPI + cell nucleus of a single FB, pointed out by the white arrows. (k) Diagram depicts the mean ± SD length/width ratio of FBs in CTRL and hydrogels ( n = 5). (l) Diagram depicts the mean ± SD nuclei size of FBs in μm 2 ( n = 5). (m) Diagram depicts the mean ± SD roundness of FB nuclei in CTRL and hydrogels ( n = 5). (n) Diagram shows the percentage of THY1 + cells (FBs) for each group ( n = 5). (o) Diagram depicts the mean ± SD number of THY1 + /EdU + cells (proliferating FBs) ( n = 5). (p) Diagram depicts the mean ± SD percentage of FBs negative for PI (alive FBs) ( n = 5). * p -value < 0.05, ** p -value < 0.01, *** p -value < 0.001.

Article Snippet: The synthetic, polypeptide hydrogel PuraMatrix (Corning), the basement membrane extract hydrogel Cultrex 3D Cell Culture Matrix Basement Membrane Extract Reduced Growth Factor (Trevigen), the laminin hydrogel Cultrex 3D Cell Culture Matrix Laminin I, the collagen hydrogel Cultrex 3D Cell Culture Matrix Rat Tail Collagen I, and the alginate peptide-coupled hydrogel Novatach (Novamatrix) were diluted according to the manufacturer’s recommendations.

Techniques: Cell Culture, Staining

Journal: ACS Applied Materials & Interfaces

Article Title: Systematic Comparison of Commercial Hydrogels Revealed That a Synergy of Laminin and Strain-Stiffening Promotes Directed Migration of Neural Cells

doi: 10.1021/acsami.2c20040

Figure Lengend Snippet: DRG neuron seeded in three Cultrex hydrogels and control. (a) Phase contrast micrographs of DRG neuron cultures on (a1) PDL/laminin-coated cell culture dishes (CTRL), (a2) Cultrex BME, (a3) Cultrex Laminin, and (a4) Cultrex Collagen. (b) Representative confocal micrographs of cultures stained for TUJ1 in green, S100 in red-brown, vimentin (VIME) in orange, and DAPI in blue. Images in the left corner are representative micrographs of a DAPI + cell nucleus of a single DRG neuron, pointed out by the white arrows. White arrowheads show SC/neurite alignment. (c) Diagram depicts the mean ± SD percentage of S100 (SC) and TUJ1 (DRG neurons) overlap ( n = 3). (d) Diagram depicts the mean ± SD nuclei size of DRG neurons in μm 2 ( n = 3). (e) Diagram depicts the mean ± SD nuclei roundness ( n = 3). (f) Diagram depicts the mean ± SD number of primary neurites ( n = 3). (g) Diagram depicts the mean ± SD number of BPs per 500 μm ( n = 3). (h) Diagram depicts the mean ± SD neurite thickness in μm ( n = 3). * p -value < 0.05, ** p -value < 0.01, *** p -value < 0.001.

Article Snippet: The synthetic, polypeptide hydrogel PuraMatrix (Corning), the basement membrane extract hydrogel Cultrex 3D Cell Culture Matrix Basement Membrane Extract Reduced Growth Factor (Trevigen), the laminin hydrogel Cultrex 3D Cell Culture Matrix Laminin I, the collagen hydrogel Cultrex 3D Cell Culture Matrix Rat Tail Collagen I, and the alginate peptide-coupled hydrogel Novatach (Novamatrix) were diluted according to the manufacturer’s recommendations.

Techniques: Control, Cell Culture, Staining

Journal: ACS Applied Materials & Interfaces

Article Title: Systematic Comparison of Commercial Hydrogels Revealed That a Synergy of Laminin and Strain-Stiffening Promotes Directed Migration of Neural Cells

doi: 10.1021/acsami.2c20040

Figure Lengend Snippet: Morphological analyses of SCs and FBs seeded on three Cultrex hydrogels and in CTRL. (a) Representative confocal micrographs of SC cultures on (a1) PLL/laminin coated cell culture dishes (CTRL), (a2) Cultrex BME, (a3) Cultrex Laminin, and (a4) Cultrex Collagen stained for S100 in green, vimentin (VIME) in gray, EdU in red, and DAPI in blue. Small images in the right corner are representative micrographs of a DAPI + cell nucleus of a single SC, pointed out by the white arrows. (b) Diagram depicts the mean ± SD length/width ratio of SCs on PLL/laminin (CTRL) and hydrogels ( n = 4). (c) Diagram depicts the mean ± SD nuclei size of SCs in μm 2 for the four groups ( n = 4). (d) Diagram depicts the roundness of SC nuclei on PLL/laminin and hydrogels ± SD ( n = 4). (e) Diagram shows the mean ± SD percentage of S100 + /EdU + cells (proliferating SCs) for each group ( n = 4). (f) Representative confocal micrographs of FB cultures on (f1) uncoated cell culture wells (CTRL), (f2) Cultrex BME, (f3) Cultrex Laminin, and (f4) Cultrex Collagen stained for THY1 in cyan, actin in gold, EdU in red, and DAPI in gray. Small images in the right corner of the image are representative micrographs of a DAPI + cell nucleus of a single FB, pointed out by the white arrows. (g) Diagram depicts the mean ± SD length/width ratio of FBs in control and hydrogels ( n = 4). (h) Diagram depicts the mean ± SD nuclei size of FBs in μm 2 ( n = 4). (i) Diagram depicts the mean ± SD roundness of FB nuclei in control and hydrogels ( n = 4). (j) Diagram depicts the mean ± SD number of THY1 + /EdU + FBs ( n = 4). * p -value < 0.05, ** p -value < 0.01, *** p -value < 0.001.

Article Snippet: The synthetic, polypeptide hydrogel PuraMatrix (Corning), the basement membrane extract hydrogel Cultrex 3D Cell Culture Matrix Basement Membrane Extract Reduced Growth Factor (Trevigen), the laminin hydrogel Cultrex 3D Cell Culture Matrix Laminin I, the collagen hydrogel Cultrex 3D Cell Culture Matrix Rat Tail Collagen I, and the alginate peptide-coupled hydrogel Novatach (Novamatrix) were diluted according to the manufacturer’s recommendations.

Techniques: Cell Culture, Staining, Control

Journal: ACS Applied Materials & Interfaces

Article Title: Systematic Comparison of Commercial Hydrogels Revealed That a Synergy of Laminin and Strain-Stiffening Promotes Directed Migration of Neural Cells

doi: 10.1021/acsami.2c20040

Figure Lengend Snippet: Live cell imaging of SCs and FBs seeded on three Cultrex hydrogels and in control. (a) Magnifications of representative images of SCs on (a1) PLL/laminin-coated cell culture dishes (CTRL), (a2) Cultrex BME, (a3) Cultrex Laminin, and (a4) Cultrex Collagen after 17 h. The colored lines each represent an SC’s migratory tracks. (b) Colored line in the coordinate system represents each cell starting at 0 (center) for each condition. (c) Diagram depicts the mean ± SD accumulated velocity (velocity accum ) of SCs in μm/min ( n = 6). (d) Diagram depicts the mean ± SD accumulated (total) distance (distance accum ) of SCs in μm ( n = 6). (e) Diagram depicts the mean ± SD effective velocity (velocity euclid ) of SCs in μm/min ( n = 6). (f) Diagram depicts the mean ± SD effective distance (distance euclid ) of SCs in μm ( n = 6). (g) Magnifications of representative images of FBs on (f1) uncoated cell culture dishes (CTRL), (f2) Cultrex BME, (f3) Cultrex Laminin, and (f4) Cultrex Collagen after 17 h. The colored lines each represent a FB’s migratory tracks. (h) Colored line in the coordinate system represents each cell starting at 0 (center) for each condition. (i) Diagram depicts the mean ± SD accumulated velocity (velocity accum ) of FBs in μm/min ( n = 4). (j) Diagram depicts the mean ± SD accumulated (total) distance (distance accum ) of FBs in μm ( n = 4). (k) Diagram depicts the mean ± SD effective velocity (velocity euclid ) of FBs in μm/min ( n = 4). (l) Diagram depicts the mean ± SD effective distance (distance euclid ) of FBs in μm ( n = 4). The bar represents the mean of all donors; * p -value < 0.05, ** p -value < 0.01, *** p -value < 0.001.

Article Snippet: The synthetic, polypeptide hydrogel PuraMatrix (Corning), the basement membrane extract hydrogel Cultrex 3D Cell Culture Matrix Basement Membrane Extract Reduced Growth Factor (Trevigen), the laminin hydrogel Cultrex 3D Cell Culture Matrix Laminin I, the collagen hydrogel Cultrex 3D Cell Culture Matrix Rat Tail Collagen I, and the alginate peptide-coupled hydrogel Novatach (Novamatrix) were diluted according to the manufacturer’s recommendations.

Techniques: Live Cell Imaging, Control, Cell Culture

Journal: ACS Applied Materials & Interfaces

Article Title: Systematic Comparison of Commercial Hydrogels Revealed That a Synergy of Laminin and Strain-Stiffening Promotes Directed Migration of Neural Cells

doi: 10.1021/acsami.2c20040

Figure Lengend Snippet: Evaluation of surface topography of hydrogels. SEM visualized smoother surface topography for (a–c left images) control coatings (PLL/laminin for SCs, uncoated for FBs, and PDL/laminin for DRGs) and (f, left image) Novatach in comparison to the features visible on (d, left image) PuraMatrix, (e, left image) Cultrex BME, and (g,h, left images) Cultrex Laminin and Cultrex Collagen. (a–e) Atomic force micrographs (a–h, right images) visualized differences in (i) surface roughness and (j) surface area between the hydrogels. (i) Diagram depicts the mean RMS roughness in nm for the control coatings and each hydrogel. (j) Diagram depicts the mean surface area for the control coatings and each hydrogel. Data are presented as mean ± SD from three technical replicates.

Article Snippet: The synthetic, polypeptide hydrogel PuraMatrix (Corning), the basement membrane extract hydrogel Cultrex 3D Cell Culture Matrix Basement Membrane Extract Reduced Growth Factor (Trevigen), the laminin hydrogel Cultrex 3D Cell Culture Matrix Laminin I, the collagen hydrogel Cultrex 3D Cell Culture Matrix Rat Tail Collagen I, and the alginate peptide-coupled hydrogel Novatach (Novamatrix) were diluted according to the manufacturer’s recommendations.

Techniques: Control, Comparison

Journal: ACS Applied Materials & Interfaces

Article Title: Systematic Comparison of Commercial Hydrogels Revealed That a Synergy of Laminin and Strain-Stiffening Promotes Directed Migration of Neural Cells

doi: 10.1021/acsami.2c20040

Figure Lengend Snippet: DRG neuron cultures seeded in hydrogels and CTRL. (a) Phase contrast micrographs of DRG neuron cultures on (1) PDL/laminin-coated cell culture dishes (CTRL) and in hydrogels (2) PuraMatrix, (3) Cultrex BME, and (4) Novatach. (b) Representative confocal micrographs of cultures stained for TUJ1 in green, S100 in red, vimentin (VIME) in orange, and DAPI in blue. Images in the left corner are representative micrographs of DAPI + cell nuclei of a single DRG neuron, indicated by white arrows. Enlargements of the neurite networks show TUJ1 and S100 stainings to assess neurite/SC co-localization indicated by white arrowheads. (c) Diagram depicts the mean ± SD percentage of S100 (SC) and TUJ1 (DRG neurons) overlap ( n = 3). (d) Diagram depicts the mean ± SD nuclei size of DRG neurons in μm 2 for the four conditions ( n = 3). (e) Diagram depicts the mean ± SD nuclei roundness ( n = 3). (f) Diagram depicts the mean ± SD number of primary neurites ( n = 3). (g) Diagram depicts the mean ± SD number of BPs per 500 μm neurite length ( n = 3). (h) Diagram depicts the mean ± SD neurite thickness in μm ( n = 3). * p -value < 0.05, ** p -value < 0.01, *** p -value < 0.001.

Article Snippet: The synthetic, polypeptide hydrogel PuraMatrix (Corning), the basement membrane extract hydrogel Cultrex 3D Cell Culture Matrix Basement Membrane Extract Reduced Growth Factor (Trevigen), the laminin hydrogel Cultrex 3D Cell Culture Matrix Laminin I, the collagen hydrogel Cultrex 3D Cell Culture Matrix Rat Tail Collagen I, and the alginate peptide-coupled hydrogel Novatach (Novamatrix) were diluted according to the manufacturer’s recommendations.

Techniques: Cell Culture, Staining

Journal: ACS Applied Materials & Interfaces

Article Title: Systematic Comparison of Commercial Hydrogels Revealed That a Synergy of Laminin and Strain-Stiffening Promotes Directed Migration of Neural Cells

doi: 10.1021/acsami.2c20040

Figure Lengend Snippet: Evaluation of SC and FB morphology, proliferation, and viability. (a) Representative phase contrast micrographs of SC cultures on (a1) poly- l -lysine (PLL)/laminin-coated cell culture dishes (CTRL), (a2) PuraMatrix, (a3) Cultrex BME, and (a4) Novatach. Hydrogel residues of PuraMatrix were free-floating in the medium [arrowheads in (a2)]. (b) Representative confocal micrographs of SCs stained for S100 in green, VIME in gray, EdU in red, and DAPI in blue. The arrowheads indicate proliferating SCs. Small images in the right corner are representative micrographs of a DAPI + cell nucleus of a single SC, indicated by white arrows. (c) Diagram depicts the mean ± SD length/width ratio of SCs on PLL/laminin (CTRL) and hydrogels ( n = 7). (d) Diagram depicts the mean ± SD nuclei size of SCs in μm 2 for the four groups ( n = 7). (e) Diagram depicts the roundness of SC nuclei on PLL/laminin and hydrogels ± SD ( n = 7). (f) Diagram shows the mean ± SD percentage of S100 + cells (SCs) for each group ( n = 6). (g) Diagram depicts the mean ± SD percentage of S100 + /EdU + cells (proliferating SCs) in each condition ( n = 7). (h) Diagram depicts the percentage of SCs negative for propidium iodide (PI) in each condition ± SD ( n = 6). (i) Representative phase contrast micrographs of FB cultures on (i1) uncoated cell culture wells (CTRL), (i2) PuraMatrix, (i3) Cultrex BME, and (i4) Novatach. (j) Representative confocal micrographs of FBs stained for THY1 in cyan, NGFR in pink, EdU in red, and DAPI in gray. The arrowhead indicates proliferating FBs, and the pink arrows indicate NGFR + SCs. Small images in the right corner of the image are representative micrographs of a DAPI + cell nucleus of a single FB, pointed out by the white arrows. (k) Diagram depicts the mean ± SD length/width ratio of FBs in CTRL and hydrogels ( n = 5). (l) Diagram depicts the mean ± SD nuclei size of FBs in μm 2 ( n = 5). (m) Diagram depicts the mean ± SD roundness of FB nuclei in CTRL and hydrogels ( n = 5). (n) Diagram shows the percentage of THY1 + cells (FBs) for each group ( n = 5). (o) Diagram depicts the mean ± SD number of THY1 + /EdU + cells (proliferating FBs) ( n = 5). (p) Diagram depicts the mean ± SD percentage of FBs negative for PI (alive FBs) ( n = 5). * p -value < 0.05, ** p -value < 0.01, *** p -value < 0.001.

Article Snippet: The synthetic, polypeptide hydrogel PuraMatrix (Corning), the basement membrane extract hydrogel Cultrex 3D Cell Culture Matrix Basement Membrane Extract Reduced Growth Factor (Trevigen), the laminin hydrogel Cultrex 3D Cell Culture Matrix Laminin I, the collagen hydrogel Cultrex 3D Cell Culture Matrix Rat Tail Collagen I, and the alginate peptide-coupled hydrogel Novatach (Novamatrix) were diluted according to the manufacturer’s recommendations.

Techniques: Cell Culture, Staining

Journal: ACS Applied Materials & Interfaces

Article Title: Systematic Comparison of Commercial Hydrogels Revealed That a Synergy of Laminin and Strain-Stiffening Promotes Directed Migration of Neural Cells

doi: 10.1021/acsami.2c20040

Figure Lengend Snippet: DRG neuron seeded in three Cultrex hydrogels and control. (a) Phase contrast micrographs of DRG neuron cultures on (a1) PDL/laminin-coated cell culture dishes (CTRL), (a2) Cultrex BME, (a3) Cultrex Laminin, and (a4) Cultrex Collagen. (b) Representative confocal micrographs of cultures stained for TUJ1 in green, S100 in red-brown, vimentin (VIME) in orange, and DAPI in blue. Images in the left corner are representative micrographs of a DAPI + cell nucleus of a single DRG neuron, pointed out by the white arrows. White arrowheads show SC/neurite alignment. (c) Diagram depicts the mean ± SD percentage of S100 (SC) and TUJ1 (DRG neurons) overlap ( n = 3). (d) Diagram depicts the mean ± SD nuclei size of DRG neurons in μm 2 ( n = 3). (e) Diagram depicts the mean ± SD nuclei roundness ( n = 3). (f) Diagram depicts the mean ± SD number of primary neurites ( n = 3). (g) Diagram depicts the mean ± SD number of BPs per 500 μm ( n = 3). (h) Diagram depicts the mean ± SD neurite thickness in μm ( n = 3). * p -value < 0.05, ** p -value < 0.01, *** p -value < 0.001.

Article Snippet: The synthetic, polypeptide hydrogel PuraMatrix (Corning), the basement membrane extract hydrogel Cultrex 3D Cell Culture Matrix Basement Membrane Extract Reduced Growth Factor (Trevigen), the laminin hydrogel Cultrex 3D Cell Culture Matrix Laminin I, the collagen hydrogel Cultrex 3D Cell Culture Matrix Rat Tail Collagen I, and the alginate peptide-coupled hydrogel Novatach (Novamatrix) were diluted according to the manufacturer’s recommendations.

Techniques: Control, Cell Culture, Staining

Journal: ACS Applied Materials & Interfaces

Article Title: Systematic Comparison of Commercial Hydrogels Revealed That a Synergy of Laminin and Strain-Stiffening Promotes Directed Migration of Neural Cells

doi: 10.1021/acsami.2c20040

Figure Lengend Snippet: Morphological analyses of SCs and FBs seeded on three Cultrex hydrogels and in CTRL. (a) Representative confocal micrographs of SC cultures on (a1) PLL/laminin coated cell culture dishes (CTRL), (a2) Cultrex BME, (a3) Cultrex Laminin, and (a4) Cultrex Collagen stained for S100 in green, vimentin (VIME) in gray, EdU in red, and DAPI in blue. Small images in the right corner are representative micrographs of a DAPI + cell nucleus of a single SC, pointed out by the white arrows. (b) Diagram depicts the mean ± SD length/width ratio of SCs on PLL/laminin (CTRL) and hydrogels ( n = 4). (c) Diagram depicts the mean ± SD nuclei size of SCs in μm 2 for the four groups ( n = 4). (d) Diagram depicts the roundness of SC nuclei on PLL/laminin and hydrogels ± SD ( n = 4). (e) Diagram shows the mean ± SD percentage of S100 + /EdU + cells (proliferating SCs) for each group ( n = 4). (f) Representative confocal micrographs of FB cultures on (f1) uncoated cell culture wells (CTRL), (f2) Cultrex BME, (f3) Cultrex Laminin, and (f4) Cultrex Collagen stained for THY1 in cyan, actin in gold, EdU in red, and DAPI in gray. Small images in the right corner of the image are representative micrographs of a DAPI + cell nucleus of a single FB, pointed out by the white arrows. (g) Diagram depicts the mean ± SD length/width ratio of FBs in control and hydrogels ( n = 4). (h) Diagram depicts the mean ± SD nuclei size of FBs in μm 2 ( n = 4). (i) Diagram depicts the mean ± SD roundness of FB nuclei in control and hydrogels ( n = 4). (j) Diagram depicts the mean ± SD number of THY1 + /EdU + FBs ( n = 4). * p -value < 0.05, ** p -value < 0.01, *** p -value < 0.001.

Article Snippet: The synthetic, polypeptide hydrogel PuraMatrix (Corning), the basement membrane extract hydrogel Cultrex 3D Cell Culture Matrix Basement Membrane Extract Reduced Growth Factor (Trevigen), the laminin hydrogel Cultrex 3D Cell Culture Matrix Laminin I, the collagen hydrogel Cultrex 3D Cell Culture Matrix Rat Tail Collagen I, and the alginate peptide-coupled hydrogel Novatach (Novamatrix) were diluted according to the manufacturer’s recommendations.

Techniques: Cell Culture, Staining, Control

Journal: ACS Applied Materials & Interfaces

Article Title: Systematic Comparison of Commercial Hydrogels Revealed That a Synergy of Laminin and Strain-Stiffening Promotes Directed Migration of Neural Cells

doi: 10.1021/acsami.2c20040

Figure Lengend Snippet: Live cell imaging of SCs and FBs seeded on three Cultrex hydrogels and in control. (a) Magnifications of representative images of SCs on (a1) PLL/laminin-coated cell culture dishes (CTRL), (a2) Cultrex BME, (a3) Cultrex Laminin, and (a4) Cultrex Collagen after 17 h. The colored lines each represent an SC’s migratory tracks. (b) Colored line in the coordinate system represents each cell starting at 0 (center) for each condition. (c) Diagram depicts the mean ± SD accumulated velocity (velocity accum ) of SCs in μm/min ( n = 6). (d) Diagram depicts the mean ± SD accumulated (total) distance (distance accum ) of SCs in μm ( n = 6). (e) Diagram depicts the mean ± SD effective velocity (velocity euclid ) of SCs in μm/min ( n = 6). (f) Diagram depicts the mean ± SD effective distance (distance euclid ) of SCs in μm ( n = 6). (g) Magnifications of representative images of FBs on (f1) uncoated cell culture dishes (CTRL), (f2) Cultrex BME, (f3) Cultrex Laminin, and (f4) Cultrex Collagen after 17 h. The colored lines each represent a FB’s migratory tracks. (h) Colored line in the coordinate system represents each cell starting at 0 (center) for each condition. (i) Diagram depicts the mean ± SD accumulated velocity (velocity accum ) of FBs in μm/min ( n = 4). (j) Diagram depicts the mean ± SD accumulated (total) distance (distance accum ) of FBs in μm ( n = 4). (k) Diagram depicts the mean ± SD effective velocity (velocity euclid ) of FBs in μm/min ( n = 4). (l) Diagram depicts the mean ± SD effective distance (distance euclid ) of FBs in μm ( n = 4). The bar represents the mean of all donors; * p -value < 0.05, ** p -value < 0.01, *** p -value < 0.001.

Article Snippet: The synthetic, polypeptide hydrogel PuraMatrix (Corning), the basement membrane extract hydrogel Cultrex 3D Cell Culture Matrix Basement Membrane Extract Reduced Growth Factor (Trevigen), the laminin hydrogel Cultrex 3D Cell Culture Matrix Laminin I, the collagen hydrogel Cultrex 3D Cell Culture Matrix Rat Tail Collagen I, and the alginate peptide-coupled hydrogel Novatach (Novamatrix) were diluted according to the manufacturer’s recommendations.

Techniques: Live Cell Imaging, Control, Cell Culture

Journal: ACS Applied Materials & Interfaces

Article Title: Systematic Comparison of Commercial Hydrogels Revealed That a Synergy of Laminin and Strain-Stiffening Promotes Directed Migration of Neural Cells

doi: 10.1021/acsami.2c20040

Figure Lengend Snippet: SCs and FBs seeded on increasing laminin concentration. (a) Representative confocal micrographs of SCs stained for NGFR in magenta, Actin in yellow, and DAPI in blue on (1) PLL and PLL/laminin coating with (2) 10, (3) 100, and (4) 1000 μg/mL laminin concentration. (b) Diagram depicts the mean ± SD length to width ratio ( n = 5). (c) Diagram depicts the mean ± SD velocity accum in μm/min ( n = 5). (d) Diagram depicts the mean ± SD distance accum in μm ( n = 5). (e) Diagram depicts the mean ± SD velocity euclid in μm/min ( n = 5). (f) Diagram depicts the mean ± SD distance euclid in μm ( n = 5). (g) Representative confocal micrographs of FBs stained for THY1 in cyan, actin in yellow, and DAPI in white on (1) uncoated wells and laminin coating with (2) 10, (3) 100, and (4) 1000 μg/mL laminin concentration. (h) Diagram depicts the mean ± SD length to width ratio ( n = 4). (i) Diagram depicts the mean ± SD velocity accum in μm/min ( n = 4). (j) Diagram depicts the mean ± SD distance accum in μm ( n = 4). (k) Diagram depicts the mean ± SD velocity euclid in μm/min ( n = 4). (l) Diagram depicts the mean ± SD distance euclid in μm ( n = 4); * p -value < 0.05, ** p -value < 0.01, *** p -value < 0.001.

Article Snippet: The synthetic, polypeptide hydrogel PuraMatrix (Corning), the basement membrane extract hydrogel Cultrex 3D Cell Culture Matrix Basement Membrane Extract Reduced Growth Factor (Trevigen), the laminin hydrogel Cultrex 3D Cell Culture Matrix Laminin I, the collagen hydrogel Cultrex 3D Cell Culture Matrix Rat Tail Collagen I, and the alginate peptide-coupled hydrogel Novatach (Novamatrix) were diluted according to the manufacturer’s recommendations.

Techniques: Concentration Assay, Staining

Journal: ACS Applied Materials & Interfaces

Article Title: Systematic Comparison of Commercial Hydrogels Revealed That a Synergy of Laminin and Strain-Stiffening Promotes Directed Migration of Neural Cells

doi: 10.1021/acsami.2c20040

Figure Lengend Snippet: Evaluation of surface topography of hydrogels. SEM visualized smoother surface topography for (a–c left images) control coatings (PLL/laminin for SCs, uncoated for FBs, and PDL/laminin for DRGs) and (f, left image) Novatach in comparison to the features visible on (d, left image) PuraMatrix, (e, left image) Cultrex BME, and (g,h, left images) Cultrex Laminin and Cultrex Collagen. (a–e) Atomic force micrographs (a–h, right images) visualized differences in (i) surface roughness and (j) surface area between the hydrogels. (i) Diagram depicts the mean RMS roughness in nm for the control coatings and each hydrogel. (j) Diagram depicts the mean surface area for the control coatings and each hydrogel. Data are presented as mean ± SD from three technical replicates.

Article Snippet: The synthetic, polypeptide hydrogel PuraMatrix (Corning), the basement membrane extract hydrogel Cultrex 3D Cell Culture Matrix Basement Membrane Extract Reduced Growth Factor (Trevigen), the laminin hydrogel Cultrex 3D Cell Culture Matrix Laminin I, the collagen hydrogel Cultrex 3D Cell Culture Matrix Rat Tail Collagen I, and the alginate peptide-coupled hydrogel Novatach (Novamatrix) were diluted according to the manufacturer’s recommendations.

Techniques: Control, Comparison

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Engineering exosomes for bone defect repair

doi: 10.3389/fbioe.2022.1091360

Figure Lengend Snippet: The properties of exosome-integrated biomaterials for bone defect repair.

Article Snippet: Enhancing exosomes activity , 3D matrix hydrogels , .

Techniques: Activity Assay, Polymer, Modification

Journal: Advanced Science

Article Title: From Snapshots to Development: Identifying the Gaps in the Development of Stem Cell‐based Embryo Models along the Embryonic Timeline

doi: 10.1002/advs.202004250

Figure Lengend Snippet: The current mouse embryo models with the platform in which they are cultured, the major finding using that model, and the morphological stage of mouse development they represent

Article Snippet: Asymmetric human epiblast (hESCs) , Polymeric hydrogel 3D matrix supplemented with matrigel/transwell filters [ ] , – Embedding hESCs on a synthetic hydrogel supplemented with matrigel, resembles a pluripotent epiblast with an apico‐basal polarity [ ] – A varying concentration of BMP4 determines the fate of the epiblast cells whereby a 1 ng mL −1 of uniform BMP4 supply breaks the symmetry and induces Bra+ cells – The epiblast clusters start with uniform BMP4 signaling but later establish a concentration gradient of Wnt to break the symmetry , Day 10 of a human embryo.

Techniques: Cell Culture, Suspension, Membrane, Activation Assay, Expressing, In Vivo

Journal: Advanced Science

Article Title: From Snapshots to Development: Identifying the Gaps in the Development of Stem Cell‐based Embryo Models along the Embryonic Timeline

doi: 10.1002/advs.202004250

Figure Lengend Snippet: The current human embryo models with the platform in which they are cultured, the major finding using that model, and the morphological stage of human development they represent

Article Snippet: Asymmetric human epiblast (hESCs) , Polymeric hydrogel 3D matrix supplemented with matrigel/transwell filters [ ] , – Embedding hESCs on a synthetic hydrogel supplemented with matrigel, resembles a pluripotent epiblast with an apico‐basal polarity [ ] – A varying concentration of BMP4 determines the fate of the epiblast cells whereby a 1 ng mL −1 of uniform BMP4 supply breaks the symmetry and induces Bra+ cells – The epiblast clusters start with uniform BMP4 signaling but later establish a concentration gradient of Wnt to break the symmetry , Day 10 of a human embryo.

Techniques: Cell Culture, Chromatin Immunoprecipitation

Journal: iScience

Article Title: Engineering confining microenvironment for studying cancer metastasis

doi: 10.1016/j.isci.2021.102098

Figure Lengend Snippet: Summary and comparison of different technologies for studying cancer cells under confinement

Article Snippet: Hydrogel-based 3D matrix , Studying cancer cell in vitro in relatively well-defined 3D matrices , ∗ , ∗∗ , ∗∗ , ∗ , ∗∗∗∗ , Studying protease activity during ECM invasion ( ); Studying invasiveness of cancer cells in a 3D matrix , etc. .

Techniques: Comparison, Imaging, Ex Vivo, Slice Preparation, In Vivo, In Vivo Imaging, Invasion Assay, In Vitro, Cell Culture, Migration, Activity Assay

Journal: Journal of orthopaedic research : official publication of the Orthopaedic Research Society

Article Title: Development of an In Vitro Intervertebral Disc Innervation Model to Screen Neuroinhibitory Biomaterials

doi: 10.1002/jor.24557

Figure Lengend Snippet: Schematic of experimental design and in vitro model fabrication. The structure of chondroitin‐4‐sulfate (CS‐A) (A) and chondroitin‐6‐sulfate (CS‐C) (B) are shown with the sulfate group in red. The locations of the neonatal dorsal root ganglion (DRG) harvested for these experiments are indicated in (C). The process of hydrogel fabrication in (D) depicts the differences between making a uniform composition hydrogel for either nucleus pulposus (NP) encapsulation or for DRG embedding versus making the in vitro disc innervation model with an inner gel and outer gel to embed a DRG at the hydrogel interface. Examples of the two different types of neurite growth quantification, traced neurite length (purple and green traces, green indicating the longest trace) and radial neurite extension (yellow line), are shown in (E) with the white line on the hydrogel interface images indicating the interface between the inner and outer gels. MACS‐A, methacrylated CS‐A; MACS‐C, methacrylated CS‐C; MAHA, methacrylated hyaluronic acid; UV, ultraviolet light. [Color figure can be viewed at wileyonlinelibrary.com]

Article Snippet: Determination of Neuroinhibition in Uniform 3D Hydrogels Hydrogel mixtures were prepared as described in the Base Hydrogel Composition section with the addition of 0.75 mg/ml laminin (344600501; R&D Systems) to enhance DRG cell attachment.

Techniques: In Vitro, Encapsulation