ABSTRACT
The purpose of this chapter is to increase understanding of some of the experimental procedures used in cartilage biology and tissue engineering research. Each culture technique has advantages, disadvantages, and limitations, and the choice of which to use will depend on the type of hypothesis tested. Chondrocytes in two-dimensional (2D) culture undergo dedifferentiation when plated on tissue culture plastic, marked by changes in the levels of chondrocyte-specific matrix proteins and cell morphology. Important parameters that influence the success of explant culture include tissue type, explant size, freshness, donor age, anatomical location, and whether the explant contains multiple tissue types. Adult rabbit articular cartilage is about 0.4 mm thick, and chondrocytes can be isolated similarly to those of bovine cartilage. Once harvested, the articular cartilage matrix can be enzymatically digested to release the chondrocytes. Cryopreservation allows for the long-term preservation of cells. Cells stored in liquid nitrogen should be viable for a minimum of 10 years.
