Endo T, Saito T, Uchida T, Onaya T
Department of Internal Medicine, University of Yamanashi Medical School, Japan.
Acta Endocrinol (Copenh). 1988 Feb;117(2):214-8. doi: 10.1530/acta.0.1170214.
A primary culture of mammalian parafollicular cells was established from rat thyroid glands in order to investigate the effects of serotonin and somatostatin on calcitonin secretion. Minced rat thyroid glands were dissociated with collagenase and cultured in a Ham's F-12K medium supplemented with calf serum (5%), insulin (1.3 X 10(-6) mol/l), hydrocortisone (10(-8) mol/l), transferrin (6.1 X 10(-9) mol/l), and glycyl-L-histidyl-L-lysin (2.5 X 10(-8) mol/l). Immunohistochemical peroxidase-antiperoxidase method revealed that the cultured parafollicular cells were immunopositive for human calcitonin, and electron microscopy demonstrated the existence of dense secretory granules in the cultured parafollicular cells. Addition of the Ca2+ to the culture medium stimulated calcitonin secretion from the cells dose-dependently as measured by radioimmunoassay. Pre-incubation of serotonin with the cells produced higher calcitonin levels in a dose-dependent manner. On the other hand, pre-incubation of somatostatin with the cells significantly inhibited calcitonin secretion.
为了研究血清素和生长抑素对降钙素分泌的影响,从大鼠甲状腺建立了哺乳动物滤泡旁细胞的原代培养物。将切碎的大鼠甲状腺用胶原酶解离,并在补充有小牛血清(5%)、胰岛素(1.3×10⁻⁶mol/L)、氢化可的松(10⁻⁸mol/L)、转铁蛋白(6.1×10⁻⁹mol/L)和甘氨酰-L-组氨酰-L-赖氨酸(2.5×10⁻⁸mol/L)的Ham's F-12K培养基中培养。免疫组织化学过氧化物酶-抗过氧化物酶方法显示,培养的滤泡旁细胞对人降钙素免疫阳性,电子显微镜显示培养的滤泡旁细胞中存在致密分泌颗粒。通过放射免疫测定法测量,向培养基中添加Ca²⁺可剂量依赖性地刺激细胞分泌降钙素。血清素与细胞预孵育以剂量依赖性方式产生更高的降钙素水平。另一方面,生长抑素与细胞预孵育可显著抑制降钙素分泌。