BIOCHEMICAL CHARACTERIZATION WITH PARATHORMONE AND CALCITONIN OF ISOLATED BONE-CELLS - PROVISIONAL IDENTIFICATION OF OSTEOCLASTS AND OSTEOBLASTS
BIOCHEMICAL CHARACTERIZATION WITH PARATHORMONE AND CALCITONIN OF ISOLATED BONE-CELLS - PROVISIONAL IDENTIFICATION OF OSTEOCLASTS AND OSTEOBLASTS
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DOI：
10.1210/endo-99-2-526
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发表时间：
1976-01-01
期刊：
影响因子：
4.8
通讯作者：
COHN, DV
中科院分区：
文献类型：
--
作者：
LUBEN, RA;WONG, GL;COHN, DV
通过使用胶原酶和胰蛋白酶的混合物进行重复消化过程，从小鼠颅骨中分离出两种代谢不同类型的骨细胞群。当用甲状旁腺素或降钙素处理时，消化早期释放的细胞显示c[环]AMP增加约2倍。这些人群被标记为 CT 类型。后来洗脱的细胞显示甲状旁腺素诱导的cAMP 增加更大，但对降钙素没有反应。这些人群被称为 PT 型。在两类人群中测量了六种代谢和酶活性：酸性和碱性磷酸酶、透明质酸合成、柠檬酸脱羧、脯氨酰羟化酶和一般蛋白质合成。尽管这两种活性均存在于两种细胞类型中，但 CT 细胞中酸性磷酸酶和透明质酸合成的基础水平较高，而 PT 细胞中碱性磷酸酶、柠檬酸脱羧酶和脯氨酰羟化酶的基础水平较高。甲状旁腺素仅在 CT 细胞中刺激酸性磷酸酶和透明质酸合成 100-200%；它仅在 PT 细胞中抑制碱性磷酸酶、柠檬酸脱羧和脯氨酰羟化酶 75-90%。单独的降钙素对除了 cAMP 产生以外的任何这些活性没有影响，但它抑制 CT 细胞中甲状旁腺素的作用。这些激素效应的敏感性、发育时间进程和强度与之前在完整颅骨中观察到的相似，表明分离的细胞系统是研究骨代谢的可靠模型。根据细胞的代谢反应，CT类型的群体可能富含破骨细胞，并且可能富含骨细胞，而PT类型的群体可能富含成骨细胞。
Two metabolically distinct types of bone cell populations were isolated from mouse calvaria by a repetitive digestive procedure with a mixture of collagenase and trypsin. Cells released early in the digestion showed approximately 2-fold increases in c[cyclic]AMP when treated with either parathormone or calcitonin. These populations were denoted CT type. Later eluting cells showed larger parathormone-induced increases in cAMP but did not respond to calcitonin. These populations were denoted PT type. Six metabolic and enzymatic activities were measured in the 2 types of populations: acid and alkaline phosphatases, hyaluronate synthesis, citrate decarboxylation, prolyl hydroxylase and general protein synthesis. Although each of these activities was present in both cell types, the basal levels of acid phosphatase and hyaluronate synthesis were higher in the CT cells, whereas alkaline phosphatase, citrate decarboxylation, and prolyl hydroxylase were higher in the PT cells. Parathormone stimulated acid phosphatase and hyaluronate synthesis by 100-200% only in the CT cells; it inhibited alkaline phosphatase, citrate decarboxylation and prolyl hydroxylase by 75-90% only in the PT cells. Calcitonin alone had no effect on any of these activities other than cAMP production, but it inhibited the action of parathormone in the CT cells. The sensitivities, time courses of development and magnitudes of these hormonal effects were similar to those observed previously in intact calvaria, indicating that the isolated cell system is a reliable model for the study of bone metabolism. Based on the metabolic responses of the cells, the CT type of populations is probably enriched in osteoclasts and, possibly, osteocytes, and the PT type of population is probably enriched in osteoblasts.