Grandolfo M, D'Andrea P, Paoletti S, Martina M, Silvestrini G, Bonucci E, Vittur F
Dipartimento di Biochimica, Biofisica e Chimica delle Macromolecole, Università di Trieste, Italy.
Calcif Tissue Int. 1993 Jan;52(1):42-8. doi: 10.1007/BF00675625.
We studied the response to culture conditions and the differentiative ability in suspension culture in alginate gels of resting chondrocytes from the preosseous cartilage of adult pig scapula. It was found that the maximum rate of chondrocyte duplication is reached at the fourth day in culture whereas the rate of proteoglycan synthesis and alkaline phosphatase expression do not gain a maximum value before the seventh day. During the culture time, the chondrocytes undergo differentiation as it is demonstrated by the alkaline phosphatase specific activity increase and by morphological criteria (hypertrophy, increase of the number of mitochondria per cell, increased endoplasmic reticulum, matrix vesicle production). The alginate gels can be easily dissolved to obtain cell populations in which the variation of cytosolic calcium concentration following a proliferative stimulus can be conveniently observed using the conventional procedure of Fura 2.
我们研究了成年猪肩胛骨骨前软骨静止软骨细胞在藻酸盐凝胶中悬浮培养时对培养条件的反应及分化能力。结果发现，软骨细胞复制的最大速率在培养的第4天达到，而蛋白聚糖合成速率和碱性磷酸酶表达在第7天之前未达到最大值。在培养期间，软骨细胞会发生分化，碱性磷酸酶比活性增加以及形态学标准（肥大、每个细胞中线粒体数量增加、内质网增多、基质小泡产生）均证明了这一点。藻酸盐凝胶可轻松溶解以获得细胞群体，使用Fura 2的常规方法可方便地观察到增殖刺激后胞质钙浓度的变化。
