X-ray microanalysis of cultured chondrocytes

Methodological aspects of X-ray microanalysis of cultured chondrocytes from rabbit auricular cartilage were investigated. The cells were grown on a Formvar-film covered carbon specimen holder with a hole, which allowed examination in STEM. Several different fluids for washing away the culture medium from the cells were used. The effect of these fluids on the elemental composition of the cells was studied. The composition of cultured chondrocytes was compared with that of auricular chondrocytes in situ. Washing with 0.3 M sucrose gave the most satisfactory results. Subsequently, the specimen holders could be frozen in liquid nitrogen and the cells freeze-dried. The effects of human growth hormone on rabbit chondrocytes were investigated by X-ray microanalysis and transmission electron microscopy. Although no ultrastructural effects of hormone treatment could be observed, marked changes in the elemental concentrations in nucleus and cytoplasm of the chondrocytes were detected.