Chondrocytes will synthesize cartilage in vitro. This protocol describes how to measure and modulate it.
- Bovine Chondrocytes (catalog # 1020 or 1021)
- X VIVO™ 15 without Phenol Red (Lonza, Walkersville, MD)
- L-ascorbic Acid (add to medium at final concentration of 50 ug/mL)
- TGFß or Other Agent to Stimulate Production, If Desired (Phenol red cannot be used if the supernatants are tested using the DMB method. See sample data on the chondrocytes page.)
- Thaw chondrocytes following recommended protocol.
- Adjust cell concentration to 1 million per mL.
- Add 100 ul to each well of a 96 well plate.
- Prepare agent to modulate cartilage synthesis in medium at twice the desired final concentration.
- Add agent to wells at 100 ul per well.
- Incubate at 37°C for 2–5 days.
- Remove supernatant and test for type II collagen or proteoglycans.