sepharose and Chondrosarcoma

Difficulty in maintaining phenotypic stability of the Swarm rat chondrosarcoma in long-term monolayer cultures has prompted investigation of alternative conditions that would enable extended maintenance of these cells, permitting use of the tumor as a model system for the long-term study of proteoglycan metabolism. Morphological analysis of the growth of the chondrosarcoma chondrocytes in agarose has shown stability of the culture over a 20 day period with respect to the ability of the cells to proliferate and synthesize an Alcian blue-positive extracellular matrix. The present study confirms these findings through analysis of the growth characteristics of the culture and the pattern of proteoglycan and collagen synthesis. The chondrocytes actively synthesize a proteoglycan-rich matrix at a rate dependent on the initial plating density and concentration of serum in the culture medium. These factors similarly affect the proliferative capabilities of the culture as demonstrated by the growth curves obtained at different culture conditions. During 20 days in culture, the cells synthesize an aggregating chondroitin sulfate proteoglycan and collagen type II, typical of cartilage and this chondrosarcoma. In addition, small molecular weight proteoglycans were found to be present at concentrations of up to 10% of the total proteoglycan population. Degradative rates are slow, the proteoglycan half-life is about 30 days, but can be enhanced with retinol, reducing the half-life to 2 days.

Topics: Animals; Cartilage; Cell Division; Cells, Cultured; Chondroitin Sulfates; Chondrosarcoma; Collagen; Cytological Techniques; Molecular Weight; Proteoglycans; Rats; Sepharose; Time Factors

Chondrocytes from the Swarm chondrosarcoma, a transplantable rat tumor, have been difficult to maintain in tissue culture for extended periods due to a time-dependent alteration of the culture to a more fibroblastic phenotype. This feature precluded the use of these cultures to examine chronic conditions that may affect cell metabolism, and the homogeneity or heterogeneity of the tumor cells within the culture population could not be examined. Use of suspension culture in agarose stabilized the chondrocyte phenotype, permitting long-term culture. Clones of tumor chondrocytes were established in agarose and were examined over 2-3 weeks for evidence that the cells were accumulating a proteoglycan-rich extracellular matrix, as determined by positive staining by Alcian blue, and were undergoing cell division. Nearly 90% of the cloned cells exhibited a prominent extracellular matrix by day 7 of culture and greater than 99% did so by day 14. Cell division did not occur to any great extent until days 6-7 of culture. After this lag, the cells appeared to undergo logarithmic growth, with a cell generation time of about 12 days. By 20 days of culture, between 80 and 90% of the initial clones contained multiple cells, indicating that nearly all the cells were in, or had entered, the cell cycle. These results suggest that the chondrocytes from the rat chondrosarcoma form a homogeneous cell population with respect to their ability to synthesize an extra-cellular matrix and divide.

Topics: Alcian Blue; Animals; Cartilage; Cell Count; Cell Separation; Cells, Cultured; Chondrosarcoma; Clone Cells; Culture Media; Extracellular Matrix; Microscopy, Electron; Rats; Rats, Inbred Strains; Sepharose; Staining and Labeling

Topics: Animals; Chondrosarcoma; Chromatography; Chromatography, DEAE-Cellulose; Electrophoresis, Polyacrylamide Gel; Female; Neoplasm Proteins; Neoplasms, Experimental; Polysaccharides; Procollagen; Rats; Sepharose; Sulfhydryl Compounds

Topics: Animals; Cells, Cultured; Chondroitin; Chondroitin Sulfates; Chondrosarcoma; Enzyme Activation; Glycosaminoglycans; Polysaccharides; Proteoglycans; Rats; Sepharose