In vitro characterization of porcine juvenile articular cartilage

Dispersed cell cultures were established from the articular cartilage of the proximal portion of the humerus of young pigs. Articular and epiphyseal portions of the cartilage were separated, minced, and enzymatically dispersed, using bacterial collagenase. Morphologically, 2 cell types were observed, using phase-contrast microscopy. Smaller polygonal cells (32.5 +/- 3.5 microns diameter) containing cytoplasmic granules were found in both areas of the cartilage. In cultures from the articular region, cells grew as monolayer cultures and initially did not demonstrate contact inhibition. In cultures from the epiphyseal region, cells grew in a multilayered manner in a colonial arrangement with cells being released from the center of the colony into the culture medium. Small granular particles (0.03 to 0.08 micron diameter) were secreted by cells in both culture systems. Particle secretion was greater in epiphyseal cultures than in articular cultures with the rate decreasing as confluency was approached. These particles stained positively for lipid and alkaline phosphatase. Acridine orange was also incorporated into the granules. The 2nd cell type, a stellate-shaped cell (60 +/- 7.6 micron diameter), was found mainly surrounding the outside of colonial areas in epiphyseal cultures. These cells did not secrete small granular particles and stained positive for factor VIII. Evaluation of cultures by scanning and transmission electron microscopy further supported the presence of 2 cell types. With scanning electron microscopy, the smaller polygonal cell was characterized by varying sizes of blebs (0.03 to 0.1 micron diameter) associated with the cell membrane and small cytoplasmic processes projecting from the cell's surface.(ABSTRACT TRUNCATED AT 250 WORDS)