Journal: Scientific Reports

Article Title: Construction of higher-order cellular microstructures by a self-wrapping co-culture strategy using a redox-responsive hydrogel

doi: 10.1038/s41598-020-63362-4

Figure Lengend Snippet: Kinetic behaviour of the detachment of a cell sheet from the redox-responsive hydrogel. ( A ) Effect of cysteine (Cys) concentration on the duration time of cell sheet detachment from the redox responsive hydrogel ( n = 3). Inset: enlarged figure after 30 min in the presence of 10–50 mM Cys. ( B ) Observation of cell sheet detachment behaviour. Hydrogels were degraded using 1–50 mM Cys. Photo (i), (ii) and (iii) present images of the cell sheet wrapping behaviour after adding 1–5, 10–20 and 30–50 mM Cys, respectively. All photos were captured by Keyence BZ-9000 microscope. Scale bar is 200 µm.

Article Snippet: Images were merged directly using the image analysis software BZ Analyzer from the Keyence BZ-9000 microscope.

Techniques: Concentration Assay, Microscopy

Journal: Scientific Reports

Article Title: Construction of higher-order cellular microstructures by a self-wrapping co-culture strategy using a redox-responsive hydrogel

doi: 10.1038/s41598-020-63362-4

Figure Lengend Snippet: Wrapping spheroids with a cell sheet by the ‘cellular Furoshiki’ technique. ( A ) Phase contrast time lapse imaging of HepG2 cells wrapped by a NIH3T3 cell sheet during the initial folding process (Vid. S2). ( B ) Representative images of the wrapped cellular structure at and after 1 h incubation. The scale bars of A and B are 500 µm. ( C ) Co-existence of HepG2 spheroids in the NIH3T3 cell sheet. Images present the double staining wrapping process between 30 min and 1 d incubation. Images in the right column are merged using the image analysis software BZ Analyzer from the Keyence BZ-9000 microscope. The scale bar is 200 µm. ( D ) Different numbers of HepG2 spheroids in the NIH3T3 cell sheet after 1 d co-culturing. 3D co-culture images were captured by the CLSM-700. The NIH3T3 cell sheet is stained with DiD red fluorescence and HepG2 spheroids are stained with Calcein-AM green fluorescence.

Article Snippet: Images were merged directly using the image analysis software BZ Analyzer from the Keyence BZ-9000 microscope.

Techniques: Imaging, Incubation, Double Staining, Software, Microscopy, Co-Culture Assay, Staining, Fluorescence

Journal: Scientific Reports

Article Title: Construction of higher-order cellular microstructures by a self-wrapping co-culture strategy using a redox-responsive hydrogel

doi: 10.1038/s41598-020-63362-4

Figure Lengend Snippet: Viability of co-cultured cells in the wrapped cellular structure. ( A ) Cell viability of HepG2 spheroids, ( B ) the NIH3T3 cell sheet and ( C ) the NIH3T3 cell sheet in the wrapped structure state after 5 d culturing. ( D ) NIH3T3-HepG2 co-culturing in the wrapped structure after 1, 3 and 5 d culturing. The initial cell numbers are 15 HepG2 spheroids, 100,000 NIH3T3 cells forming a monolayer and 50 collagen beads. Viable cells are stained green with calcein-AM and dead cells are stained red by propidium iodide. Images were merged directly using the image analysis software BZ Analyzer from the Keyence BZ-9000 microscope. Scale bar is 100 µm. ( E ) Evaluation of the cell viability ratio for different wrapped structures after 5 d culturing. Data of viable cells at day 5 were normalized to cell viability at day 1. The initial cell numbers are 15 HepG2 spheroids, 100,000 NIH3T3 cells forming a monolayer and 4000 cells/well HUVECs. Collagen microparticle numbers are approximately 50 (+), 150 (++) and 250 (+++) in the wrapped cellular structures. Error bars denote standard deviation ( n = 3). * p < 0.05, ** p < 0.01, *** p < 0.001 and **** p < 0.0001 when compared with that of the wrapped structure without collagen beads.

Article Snippet: Images were merged directly using the image analysis software BZ Analyzer from the Keyence BZ-9000 microscope.

Techniques: Cell Culture, Staining, Software, Microscopy, Standard Deviation

Journal: Scientific Reports

Article Title: Construction of higher-order cellular microstructures by a self-wrapping co-culture strategy using a redox-responsive hydrogel

doi: 10.1038/s41598-020-63362-4

Figure Lengend Snippet: Kinetic behaviour of the detachment of a cell sheet from the redox-responsive hydrogel. ( A ) Effect of cysteine (Cys) concentration on the duration time of cell sheet detachment from the redox responsive hydrogel ( n = 3). Inset: enlarged figure after 30 min in the presence of 10–50 mM Cys. ( B ) Observation of cell sheet detachment behaviour. Hydrogels were degraded using 1–50 mM Cys. Photo (i), (ii) and (iii) present images of the cell sheet wrapping behaviour after adding 1–5, 10–20 and 30–50 mM Cys, respectively. All photos were captured by Keyence BZ-9000 microscope. Scale bar is 200 µm.

Article Snippet: Images in the right column are merged using the image analysis software BZ Analyzer from the Keyence BZ-9000 microscope.

Techniques: Concentration Assay, Microscopy

Journal: Scientific Reports

Article Title: Construction of higher-order cellular microstructures by a self-wrapping co-culture strategy using a redox-responsive hydrogel

doi: 10.1038/s41598-020-63362-4

Figure Lengend Snippet: Wrapping spheroids with a cell sheet by the ‘cellular Furoshiki’ technique. ( A ) Phase contrast time lapse imaging of HepG2 cells wrapped by a NIH3T3 cell sheet during the initial folding process (Vid. S2). ( B ) Representative images of the wrapped cellular structure at and after 1 h incubation. The scale bars of A and B are 500 µm. ( C ) Co-existence of HepG2 spheroids in the NIH3T3 cell sheet. Images present the double staining wrapping process between 30 min and 1 d incubation. Images in the right column are merged using the image analysis software BZ Analyzer from the Keyence BZ-9000 microscope. The scale bar is 200 µm. ( D ) Different numbers of HepG2 spheroids in the NIH3T3 cell sheet after 1 d co-culturing. 3D co-culture images were captured by the CLSM-700. The NIH3T3 cell sheet is stained with DiD red fluorescence and HepG2 spheroids are stained with Calcein-AM green fluorescence.

Article Snippet: Images in the right column are merged using the image analysis software BZ Analyzer from the Keyence BZ-9000 microscope.

Techniques: Imaging, Incubation, Double Staining, Software, Microscopy, Co-Culture Assay, Staining, Fluorescence

Journal: Scientific Reports

Article Title: Construction of higher-order cellular microstructures by a self-wrapping co-culture strategy using a redox-responsive hydrogel

doi: 10.1038/s41598-020-63362-4

Figure Lengend Snippet: Viability of co-cultured cells in the wrapped cellular structure. ( A ) Cell viability of HepG2 spheroids, ( B ) the NIH3T3 cell sheet and ( C ) the NIH3T3 cell sheet in the wrapped structure state after 5 d culturing. ( D ) NIH3T3-HepG2 co-culturing in the wrapped structure after 1, 3 and 5 d culturing. The initial cell numbers are 15 HepG2 spheroids, 100,000 NIH3T3 cells forming a monolayer and 50 collagen beads. Viable cells are stained green with calcein-AM and dead cells are stained red by propidium iodide. Images were merged directly using the image analysis software BZ Analyzer from the Keyence BZ-9000 microscope. Scale bar is 100 µm. ( E ) Evaluation of the cell viability ratio for different wrapped structures after 5 d culturing. Data of viable cells at day 5 were normalized to cell viability at day 1. The initial cell numbers are 15 HepG2 spheroids, 100,000 NIH3T3 cells forming a monolayer and 4000 cells/well HUVECs. Collagen microparticle numbers are approximately 50 (+), 150 (++) and 250 (+++) in the wrapped cellular structures. Error bars denote standard deviation ( n = 3). * p < 0.05, ** p < 0.01, *** p < 0.001 and **** p < 0.0001 when compared with that of the wrapped structure without collagen beads.

Article Snippet: Images in the right column are merged using the image analysis software BZ Analyzer from the Keyence BZ-9000 microscope.

Techniques: Cell Culture, Staining, Software, Microscopy, Standard Deviation