Figure 5.

MSC dispersion on cartilage with increasing concentration of HA. MSCs were cultured until the second passage before use in this experiment. Cartilage discs were used to plug the bottom of a 96-well place in the correct orientation. Media formulations were then added to each well. The media formulations used were (a) 0.5 mg/mL HA media, (b) 1 mg/mL HA media, (c) 2 mg/mL HA media, (d) 3 mg/mL HA media, (e) 4 mg/mL HA media, and (f) 5 mg/mL HA media. MSCs were then stripped using TrypLE, counted using standard enumeration technique, and then seeded onto the cartilage discs at a density of 5 × 10³ cells/disc. The 96-well plate was then incubated for 24 hours at 37°C and 5% CO₂. After 24 hours the cartilage discs were washed in PBS and then fixed in 4% paraformaldehyde. Cartilage discs were washed in PBS and then stained in using Hoechst 33342 (blue) and again washed five times in PBS. Cartilage discs were then permeabilised in 0.1% Triton X-100, washed in PBS, and blocked in 1% Bovine serum albumin in PBS before being stained with F-actin-specific Alexa Fluor 488-phalloidin (green). All images were taken using the OLYMPUS FLUOVIEW FV1000 IX81 inverted confocal microscope at 10x magnification.