Separation of fractions enriched in hypertrophic cells and proliferative cells has been achieved by density gradient centrifugation of cells from collagenase digests of rabbit epiphyseal cartilage. Concentrated suspensions of cells are centrifuged on a continuous Percoll density gradient. Hypertrophic cells remain in the upper part of the gradient and proliferative zone cells move to the lower regions. The resultant fractions show differences in mean cell diameter, alkaline phosphatase activity, morphology and synthetic activity in culture. Fractions rich in hypertrophic cells contain larger cells and more alkaline phosphatase activity than those enriched in proliferative cells. In culture the hypertrophic cells flatten as large irregular polygonal cells, whereas proliferative fractions form smaller spindle-shaped cells. In micromass culture hypertrophic fractions incorporate less 35S-sulphate and 14C-proline, and less tritiated thymidine than do proliferative fractions. These results suggest a general reduction in matrix and DNA synthesis with the attainment of the fully differentiated hypertrophic state, coincident with the expression of alkaline phosphatase activity and mineralisation of the cartilage matrix.

译文

通过从兔epi骨软骨的胶原酶消化物中分离细胞的密度梯度离心,可以分离富含肥厚细胞和增殖细胞的级分。在连续的Percoll密度梯度上离心细胞的浓缩悬浮液。肥厚细胞保留在梯度的上部,增殖区细胞移动到下部区域。所得组分在培养物中显示出平均细胞直径,碱性磷酸酶活性,形态和合成活性的差异。富含肥厚细胞的组分比富含增殖细胞的组分含有更大的细胞和更多的碱性磷酸酶活性。在培养物中,肥厚细胞变平为大型不规则多边形细胞,而增殖部分形成较小的纺锤形细胞。在微质量培养中,肥厚级分与增殖级分相比,含有更少的35s硫酸盐和14c-脯氨酸,以及更少的tristiate胸苷。这些结果表明,随着完全分化的肥厚状态的实现,基质和DNA合成普遍减少,与碱性磷酸酶活性的表达和软骨基质的矿化相吻合。

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