Cell fractions from rat rib growth cartilage. Morphological and X-ray microanalytical investigation

For in vitro studies of differentiation and proliferation of chondrocytes, a need arose to prepare enriched cell fractions for analysis of the effects of different growth promoting factors. Therefore chondrocytes were isolated from rat rib growth cartilage in young rats by collagenase digestion and subsequently three cell fractions were prepared by centrifugation in a step gradient of Percoll. In a previous paper, matrix molecules synthesized by each fraction were characterized biochemically. In the present study, ultrastructure and elemental content of the fractionated chondrocytes were analyzed by scanning electron microscopy, transmission electron microscopy, and energy dispersive X-ray microanalysis. DNA-synthetic activity was measured by means of autoradiography of 3H-thymidine incorporating cells. Both morphology and elemental concentrations differed between fractions and were characteristic for each fraction. The cell fraction with the lowest density consisted of large, polygonal cells that became flattened in culture. The reduced synthetic activity and markedly lowered K content of these cells suggest that they originate from the hypertrophic zone of the growth plate. The cells in the fraction with the highest density were rather homogeneous in size and shape, and had a well developed rough endoplasmic reticulum and Golgi complex, characteristic of proliferating and resting chondrocytes. Concentrations of P and K were also significantly higher in this fraction. The fraction with intermediate density contained an admixture of cells with a predominance of proliferating cells with high DNA-synthetic activity.