human fibronectin solution  (Corning Life Sciences)

Journal: Scientific Reports

Article Title: A novel strategy to facilitate uniform epithelial cell maturation using liquid–liquid interfaces

doi: 10.1038/s41598-024-63115-7

Figure Lengend Snippet: Time course of cell density of MDCK cells cultured on fibronectin-coated or non-coated liquid–liquid or solid–liquid interfaces. As shown in Supplementary Fig. , liquid–liquid interfaces in 24-well plate were prepared under sterile conditions, immediately before the initiation of confluent culture. Viable cells were seeded at a density of 5.3 × 10 4 cells/cm 2 in 24-well plates, and the cells in each culture vessel were counted at t = 24, 48, 72, 96 h. Open circles or triangles represent fibronectin non-coated solid–liquid or liquid–liquid interfaces; closed circles or triangles represent fibronectin-coated solid–liquid or liquid–liquid interfaces. Data were obtained from three independent cultures. Statistical significance for Student’s t-test was set at * p < 0.05; or ** p < 0.01.

Article Snippet: A 20 μg/mL fibronectin solution (1918-FN-02M; R&D Systems, Minneapolis, MN, USA) in phosphate-buffered saline (PBS) was slowly introduced into the culture vessel and incubated for 1 h at room temperature (32–37 °C), and then washed four-times with PBS.

Techniques: Cell Culture, Sterility

Journal: Scientific Reports

Article Title: A novel strategy to facilitate uniform epithelial cell maturation using liquid–liquid interfaces

doi: 10.1038/s41598-024-63115-7

Figure Lengend Snippet: Immunostaining images of cell nuclei (DAPI, blue), F-actin (phalloidin, red) and tight junction protein (ZO-2, green) in MDCK cells cultured on fibronectin-coated or non-coated liquid–liquid or solid–liquid interfaces at t = 24 h and t = 72 h. Panels ( a-1 – a-4 , b-1 – b-4 , c-1 – c-4 , d-1 – d-4 ) show the enlargements of demarcated boxed areas in panels E–H. The scale bars indicate 5 mm (yellow) or 100 μm (white).

Article Snippet: A 20 μg/mL fibronectin solution (1918-FN-02M; R&D Systems, Minneapolis, MN, USA) in phosphate-buffered saline (PBS) was slowly introduced into the culture vessel and incubated for 1 h at room temperature (32–37 °C), and then washed four-times with PBS.

Techniques: Immunostaining, Cell Culture

Journal: Scientific Reports

Article Title: A novel strategy to facilitate uniform epithelial cell maturation using liquid–liquid interfaces

doi: 10.1038/s41598-024-63115-7

Figure Lengend Snippet: Frequencies of local squares, F X , against nucleus density on a local square, X LN , of MDCK cells cultured on fibronectin-coated or non-coated liquid–liquid or solid–liquid interfaces at t = 24 h ( a – d ) and t = 72 h ( e – h ). Data were obtained from 75 local squares in three independent cultures.

Article Snippet: A 20 μg/mL fibronectin solution (1918-FN-02M; R&D Systems, Minneapolis, MN, USA) in phosphate-buffered saline (PBS) was slowly introduced into the culture vessel and incubated for 1 h at room temperature (32–37 °C), and then washed four-times with PBS.

Techniques: Cell Culture

Journal: Scientific Reports

Article Title: A novel strategy to facilitate uniform epithelial cell maturation using liquid–liquid interfaces

doi: 10.1038/s41598-024-63115-7

Figure Lengend Snippet: Correlation between homogeneity index, H LN , and frequency of ZO-2-positive cells, F Z, values of MDCK cells cultured on liquid–liquid or solid–liquid interfaces at t = 72 h. Liquid–liquid interfaces in 24-well plate were prepared under sterile conditions, immediately before the initiation of confluent culture (Supplementary Fig. ). Viable cells were seeded at a density of 5.3 × 10 4 cells/cm 2 in 24-well plates. The cells cultured on fibronectin-coated or non-coated liquid–liquid or solid–liquid interfaces. In culture condition of Y27632-exposure, the cells on non-coated liquid–liquid or solid–liquid interfaces were cultured in media with and without 10 μg/mL Y27632 at t = 0–48 h. From t = 48 h, the cells were cultured in media without 10 μg/mL Y27632. The staining images of nuclei and ZO-2 at t = 72 h were analyzed using the ImageJ software. The plots show H LN and F Z values of cultures grown on solid–liquid interfaces (shaded circles) or liquid–liquid interfaces (shaded triangles) when exposed to Y27632, as well as in cultures grown on non-coated solid–liquid interfaces (an opened circle), non-coated liquid–liquid interfaces (an opened triangle), fibronectin-coated solid–liquid interfaces (a closed circle), and fibronectin-coated liquid–liquid interface (a closed triangle) at t = 72 h. Exposure of cells to Y27632 was performed at t = 0–48 h. Data were obtained from three independent experiments. Statistical significance was determined via one-way analysis of variance (ANOVA) followed by Tukey–Kramer post-hoc test (** p < 0.01; * p < 0.05).

Article Snippet: A 20 μg/mL fibronectin solution (1918-FN-02M; R&D Systems, Minneapolis, MN, USA) in phosphate-buffered saline (PBS) was slowly introduced into the culture vessel and incubated for 1 h at room temperature (32–37 °C), and then washed four-times with PBS.

Techniques: Cell Culture, Sterility, Staining, Software