Wakabayashi S, Kagawa S, Nakao K, Mimura K, Matsuoka A
J Endocrinol. 1984 Dec;103(3):377-81. doi: 10.1677/joe.0.1030377.
Monolayer cultures of the pancreas of the neonatal rat were maintained for 7 days in glucose-depleted TCM 199 medium, supplemented with 5.5 mmol galactose/l, with or without 0.1 mmol 2-deoxyglucose/l. Under culture conditions without 2-deoxyglucose, the responsiveness of B cells supported on galactose was totally abolished by day 7 of culture, and islets degenerated. In contrast, the addition of 2-deoxyglucose to the galactose-supplemented medium promoted the survival and the function of B cells even in a glucose-depleted environment and, in addition, yielded the monolayers mostly consisting of endocrine cells by destroying fibroblasts selectively. On day 7 the recovery of insulin in the cells was higher than that of the cells grown in medium with galactose alone (10.7-fold), and than the initial level at day 0 (twofold). Furthermore, the response to an acute challenge with 16.7 mmol glucose/l was 3.3-fold, to 10 mmol leucine/l it was 8.5-fold, and to 10 mmol 2-ketoisocaproate/l it was 10.9-fold above each value observed on day 1. In summary, the above data indicate that morphologically intact, functionally competent endocrine B cells can be grown in medium free from glucose.
新生大鼠胰腺的单层培养物在不含葡萄糖的TCM 199培养基中维持7天，该培养基添加有5.5 mmol/升半乳糖，添加或不添加0.1 mmol/升2-脱氧葡萄糖。在没有2-脱氧葡萄糖的培养条件下，培养至第7天时，半乳糖支持的B细胞的反应性完全丧失，胰岛退化。相比之下，在添加半乳糖的培养基中添加2-脱氧葡萄糖，即使在无葡萄糖的环境中也能促进B细胞的存活和功能，此外，通过选择性地破坏成纤维细胞，使单层培养物主要由内分泌细胞组成。在第7天时，细胞中胰岛素的恢复量高于仅在含半乳糖培养基中生长的细胞（10.7倍），也高于第0天的初始水平（两倍）。此外，对16.7 mmol/升葡萄糖的急性刺激的反应是第1天观察值的3.3倍，对10 mmol/升亮氨酸的反应是8.5倍，对10 mmol/升2-酮异己酸的反应是第1天观察值的10.9倍。总之，上述数据表明，形态完整、功能正常的内分泌B细胞可以在无葡萄糖的培养基中生长。
