Brown E M, Hurwitz S, Aurbach G D
Endocrinology. 1976 Dec;99(6):1582-8. doi: 10.1210/endo-99-6-1582.
A method is described for the preparation of isolated bovine parathyroid cells. Fresh, minced bovine parathyroid tissue is incubated with 2 mg/ml collagenase and 50 mug/ml DNAse for 1 h at 37 C with periodic pipetting. Parthyroid cells are separated from contaminating fat cells and red cells by low speed centrifugation. The resulting cell preparation is indistinguishable from parathyroid cells in intact bovine glands by light and electron microscopy. Hormone release is linear for at least 3 h at both high (2.0 mM) and low (0.5 mM) calcium concentrations and is inversely proportional to divalent cation concentration between 0.3 mM and 1.0 mM calcium as been observed previously both in vivo and in vitro. As in previous studies, hormone release is also stimulated up to 2-fold by 10(6) M (-)isoproterenol, an effect blocked by 10(-6)M (-)propranolol, suggesting a beta-adrenergic effect. Such a cell preparation should be useful for studying hormone binding and effects on cyclic nudleotides and cellular transport phenomena in both normal amd abnormal tissue.
本文描述了一种制备分离的牛甲状旁腺细胞的方法。将新鲜切碎的牛甲状旁腺组织与2mg/ml胶原酶和50μg/ml脱氧核糖核酸酶在37℃孵育1小时，期间定期进行移液操作。通过低速离心将甲状旁腺细胞与污染的脂肪细胞和红细胞分离。通过光学显微镜和电子显微镜观察，所得细胞制剂与完整牛腺体中的甲状旁腺细胞无法区分。在高钙（2.0mM）和低钙（0.5mM）浓度下，激素释放至少3小时呈线性，并且如先前在体内和体外观察到的那样，在0.3mM至1.0mM钙之间，激素释放与二价阳离子浓度成反比。与先前的研究一样，10⁶M（-）异丙肾上腺素可使激素释放增加至2倍，这种作用可被10⁻⁶M（-）普萘洛尔阻断，提示存在β-肾上腺素能效应。这种细胞制剂对于研究正常和异常组织中激素结合以及对环核苷酸和细胞转运现象的影响应该是有用的。
