Journal: ACS biomaterials science & engineering

Article Title: 3-D Human Renal Tubular Organoids Generated from Urine-Derived Stem Cells for Nephrotoxicity Screening

doi: 10.1021/acsbiomaterials.0c01468

Figure Lengend Snippet: Viability of human USCs in 3-D organoids. (a) Initially seeded at increasing numbers of cells, after 1 week culture as assessed by bright-field microscopy; (b) H&E staining; (c) live/dead cell staining and fluorescent imaging; (d) cell viability of human USC 3-D organoids at different numbers of initial seeding cells one after 1 week culture, assessed by live/dead cell imaging assay; (e) Size of the 3-D organoids derived from human USC increased proportional with the numbers of cells initially seeded after 1 week culture.

Article Snippet: Whole organoids, or sections of organoids, after 14 days culture were fixed with 4% paraformaldehyde for 15 min at room temperature and blocked with serum-free block solution (Dako, Denmark) for an additional 15 min. Four types of organoids were generated: nondifferentiated USCs (no k-ECM), differentiated USCs (with k-ECM), USCs with collagen type I (rat tail, 3.5 μ g/mL, Corning Inc, Corning, NY) or human renal cells as control.

Techniques: Microscopy, Staining, Imaging, Derivative Assay

Journal: Biomedicines

Article Title: Evaluation of the Effectiveness of an Innovative Polycomponent Formulation on Adult and Aged Human Dermal Fibroblasts

doi: 10.3390/biomedicines11092410

Figure Lengend Snippet: Effect of K formulation on type I collagen levels and synthesis in adult and aged HDFs. ( A ) Western blot assay for intracellular collagen I was performed on adult and aged HDFs treated with increasing concentrations of K for 72 h. The values obtained by densitometric analysis were normalized vs. GAPDH protein. Data are reported as the means ± SEM of two independent experiments. Images of representative immunoblots are shown. ( B ) Representative immunofluorescence images of adult and aged HDFs treated for 72 h with K (5%) and stained with anti-collagen I antibody (green). Nuclei were counterstained with DAPI (blue). The images are representative of three independent experiments in duplicate. All images were acquired at 100× magnification. ( C ) Collagen levels in fibroblasts treated as described above were evaluated in cell supernatants, using ELISA kit. Results are relative to mean values ± SEM of three experiments performed in duplicate. ( D ) P4HA1 levels were evaluated by Western blot analysis on adult and aged HDFs treated as described above. In all cases, the comparative analysis of data was performed using one-way ANOVA followed by Dunnett’s post hoc test (* p < 0.05, ** p < 0.01, *** p < 0.001).

Article Snippet: Briefly, acid-extracted type I collagen (5 mg/mL) mixture was made on ice with rat tail collagen I (Enzo Life Sciences, Lausen, Switzerland) in acetic acid 0.2%.

Techniques: Formulation, Western Blot, Immunofluorescence, Staining, Enzyme-linked Immunosorbent Assay