Journal: Aging Cell
Article Title: Cellular senescence contributes to spontaneous repair of the rat meniscus
doi: 10.1111/acel.14385
Figure Lengend Snippet: Effect of senescence‐associated secretory phenotype (SASP) factors derived from senescent rat synovial fibroblasts (SFs). (a) Schema of senescence induction in rat SFs using IL‐1β and TNF‐α (IL/TNF). (b) Phase contrast and brightfield images of senescence‐associated beta‐galactosidase (SA‐β‐gal) staining. The percentage of SA‐β‐gal–positive cells is shown. (c) Scheme of the cell proliferation assay. Rat SFs were cultured in conditioned medium collected from the control and IL/TNF SFs (cont‐CM and IL/TNF‐CM) for 2, 4, and 6 days. (d) Representative images of cells stained with crystal violet. (e) Relative cell numbers at days 0, 2, 4, and 6. (f) Scheme of chondrogenic differentiation assay. Rat SFs were pretreated with cont‐CM and IL/TNF‐CM for 1 day and then subjected to 3D chondrogenic culture. (g) Representative images of safranin O staining of chondrogenic spheroids. (h) Quantification of the glycosaminoglycan (GAG)/DNA ratio. (i) Relative gene expression of SOX9, ACAN, COL2A1, COL1A1, and COL10A1. * p < 0.05, ** p < 0.01, *** p < 0.005, **** p < 0.0005. Each experiment was performed in triplicate and repeated twice. Representative data are presented.
Article Snippet: Normal rat SFs were induced to senesce by culturing for 5 days in growth medium with 1 ng/mL recombinant rat IL‐1β and TNF‐α (IL/TNF; Peprotech, Rocky Hill, NJ, USA).
Techniques: Derivative Assay, Staining, Proliferation Assay, Cell Culture, Control, Differentiation Assay, Gene Expression