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  • Title: [Quantitative analysis of nuclear DNA of rat chondrocytes during the course of growth and aging--Feulgen-DNA cytofluorometry method].
    Author: Kusuzaki K, Yamashita F, Sakakida K, Kamachi M, Ashihara T.
    Journal: Nihon Seikeigeka Gakkai Zasshi; 1983 Jul; 57(7):721-7. PubMed ID: 6655320.
    Abstract:
    There have been no reported studies on the Feulgen-DNA cytofluorometry of the cartilage cells. We have attempted to devise a method of cell separation from the epiphyseal and articular cartilages of the rats, and to analyze by cytofluorometry the changes in the ploidy patterns of these chondrocytes during growth and ageing of the animals. Chondrocytes were isolated from the proximal cartilage of tibia by dual enzymatic digestions of the cartilage matrix with papain and collagenase, followed by mechanical cell separation with scissors and a micro-homogenizer, and were smeared onto the object glass with PBS. These procedures were found to be suitable for the Feulgen-DNA cytofluorometry of the chondrocytes from our repeated studies. We also carried out Feulgen-DNA cytofluorometry combined with 3H-thymidine autoradiography to determine cellular DNA content of the DNA synthetic chondrocytes in the epiphyseal cartilage. It has been clarified that during the growth course of the rats, the chondrocytes of the epiphyseal cartilage consist of many mononuclear diploid cells, a few mononuclear tetraploid cells and of some fraction of the cells having intermediate DNA values between the diploid and tetraploid levels. Those cells with intermediate DNA values, after autoradiographic studies, were found to correspond to DNA synthetic cells, indicating cell proliferative activity. It has been shown that during ageing of the rats, most of the chondrocytes from the articular cartilage are mononuclear diploid cells. The distribution of each cellular DNA content at the diploid level as determined by Feulgen-DNA cytofluorometry was shown to become gradually broader.
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