Kagawa S, Wakabayashi S, Nakao K, Matsuoka A
Department of Clinical Pathology, Hyogo College of Medicine, Nishinomiya, Japan.
Life Sci. 1987 Nov 9;41(19):2241-9. doi: 10.1016/0024-3205(87)90522-4.
Monolayer cultures of pancreatic B cells of 3-week-old rats were kept for 7 days in medium with 5.5 mM glucose plus 1 mM 2-deoxy-2-fluoroglucose or for 4 days in medium with 5.5 mM glucose alone, following exposure for 3 days to a medium with 5.5 mM glucose plus 5 microM iodoacetic acid. Addition of the deoxyglucose or iodoacetic acid caused a selective deletion of fibroblasts, yielding large clusters that consisted mostly of islet cells. At the early stage of culture in medium with 16.7 mM glucose (day 4), the response of B cells to 16.7 mM glucose included only a small rise in insulin secreted during the first and second phases, and that to 10 mM of leucine and 2-ketoisocaproate was monophasic. After culturing for 7 days, these three secretagogues markedly stimulated insulin secretion by B cells cultured in both media, with a significant rise in secondary phase secretion. However, quantitative relationships differed. Thus, the response (total insulin secreted during a 30-min stimulation) of B cells in 2-deoxy-2-fluoroglucose to glucose was 155%, to leucine 185% and 2-ketoisocaproate 126% of that of cells exposed to iodoacetic acid. In conclusion, the present results suggest that B cells of 3-week-old rat may be immature, and that medium containing 2-deoxy-2-fluoroglucose is beneficial to continued maturation of the response in vitro.
将3周龄大鼠的胰腺B细胞单层培养物在含5.5 mM葡萄糖加1 mM 2-脱氧-2-氟葡萄糖的培养基中培养7天，或在仅含5.5 mM葡萄糖的培养基中培养4天，此前这些细胞已在含5.5 mM葡萄糖加5 microM碘乙酸的培养基中暴露3天。添加脱氧葡萄糖或碘乙酸会导致成纤维细胞选择性缺失，形成主要由胰岛细胞组成的大细胞簇。在含16.7 mM葡萄糖的培养基中培养的早期阶段（第4天），B细胞对16.7 mM葡萄糖的反应仅包括在第一和第二阶段分泌的胰岛素有小幅升高，对10 mM亮氨酸和2-酮异己酸的反应是单相的。培养7天后，这三种促分泌剂显著刺激了在两种培养基中培养的B细胞的胰岛素分泌，第二阶段分泌有显著增加。然而，定量关系有所不同。因此，在2-脱氧-2-氟葡萄糖中培养的B细胞对葡萄糖的反应（30分钟刺激期间分泌的总胰岛素量）是暴露于碘乙酸的细胞的155%，对亮氨酸是185%，对2-酮异己酸是126%。总之，目前的结果表明3周龄大鼠的B细胞可能不成熟，并且含2-脱氧-2-氟葡萄糖的培养基有利于体外反应的持续成熟。
