Countercurrent centrifugal elutriation. High-resolution method for the separation of growth-plate chondrocytes

Countercurrent centrifugal elutriation was used to separate, on the basis of size, isolated growth-plate chondrocytes in chicks. The mean cellular volume, activity of alkaline phosphatase, and synthesis of type-X collagen increased progressively in each of seven successive fractions. Slices of tissues that contained either proliferating or hypertrophic chondrocytes were also removed by manual dissection from the superficial and deep regions of the growth plate. These cells demonstrated differences in size and biochemistry that were similar to those observed in chondrocytes that were separated by elutriation. These differences included increased synthesis of proteoglycan and collagen in the larger chondrocytes. Radiolabeled hypertrophic chondrocytes were mixed with unlabeled resting and proliferating chondrocytes, and then were separated by elutriation. The radioactivity was recovered in fractions that contained the largest cells, confirming that differences in the sizes of the cells can be used to effect a zonal separation by centrifugal elutriation.