Abstract:

BACKGROUND: Cartilage cells are target cells during cartilage damage, which are the main cells secreted inflammatory cytokines. It is difficult to isolate osteoarthritis cartilage cells.
OBJECTIVE: To isolate and culture rabbit articular cartilage cells and to observe the morphological characteristics of rabbit cartilage cells.
METHODS: Articular cartilage cells were obtained by mechanical-enzymatic digestion from the cartilage of 4-week-old New Zealand white rabbits. The cells were identified by phase-contrast microscope, growth curve, hematoxylin-eosin staining, Alician blue staining and type Ⅱ collagen immunofluorescence staining.
RESULTS AND CONCLUSION: The cultivation of cartilage cells presented with polygonal or triangle, and turned to dedifferentiation after third generation. Morphology and immunofluorescence staining showed that stable phenotype could be maintained within three generations. 90% cartilage cells maintained polygonal or triangle with circular or oval nuclei. Hematoxylin-eosin staining showed nucleus stained purple, cartilage matrix stained red, and surrounding cells or cytoplasm stained amaranth. Alician blue staining showed that cell plasma and membrane presented with dark blue. Type Ⅱ collagen immunofluorescence staining showed clear cytoplasm and cell membrane, which presented with green fluorescence, but there was not green fluorescence in the nucleus. Experimental results demonstrated that, a system that can isolate and culture cartilage cell is successful established. Over 90% cartilage cells grow well within three generations.