ESTABLISHMENT OF 2-MERCAPTOETHANOL-DEPENDENT DIFFERENTIATED INSULIN-SECRETING CELL-LINES
ESTABLISHMENT OF 2-MERCAPTOETHANOL-DEPENDENT DIFFERENTIATED INSULIN-SECRETING CELL-LINES
复制标题
DOI：
10.1210/en.130.1.167
复制
发表时间：
1992-01-01
期刊：
影响因子：
4.8
通讯作者：
WOLLHEIM, CB
中科院分区：
文献类型：
--
作者：
ASFARI, M;JANJIC, D;WOLLHEIM, CB
新的胰岛素分泌细胞系（INS-1 和 INS-2）是从 X 射线诱导的大鼠可移植胰岛素瘤中分离的细胞建立的。 发现这些细胞的持续生长依赖于还原剂2-巯基乙醇。 去除这种硫醇化合物导致细胞总谷胱甘肽水平下降 15 倍。 这些细胞增殖缓慢（群体倍增时间约为 100 小时），并且通常表现出天然 β 细胞的典型形态特征。 大多数细胞的胰岛素染色呈阳性，并且不与针对其他胰岛激素的抗体发生反应。 免疫反应性胰岛素的含量约为8-mu-g/10(6)细胞，相当于天然β细胞含量的20%。 这些细胞合成胰岛素原 I 和 II，并表现出与大鼠胰岛中观察到的相似的两种前体激素的转化率。 然而，葡萄糖未能刺激胰岛素原生物合成的速率。 在静态培养中，葡萄糖刺激漂浮细胞簇或附着细胞分泌胰岛素。 在灌注条件下，10 mM 而不是 1 mM 葡萄糖使分泌增强 2.2 倍。 在毛喉素和 3-异丁基-1-甲基黄嘌呤存在下，葡萄糖浓度从 2.8-20 mM 增加导致分泌速率增加 4 倍。 葡萄糖还使 INS-1 细胞去极化并提高胞质 Ca2+ 浓度。 这表明葡萄糖仍然能够在质膜上引发部分离子事件，从而导致胰岛素分泌。 INS-1 细胞的结构和功能特征在 2 年时间内（约 80 代）保持不变。 尽管 INS-2 细胞尚未完全表征，但它们的胰岛素含量与 INS-1 细胞相似，并且它们仍然对作为促分泌剂的葡萄糖部分敏感。 INS-1 细胞保留了 β 细胞表面抗原，如与抗神经节苷脂单克隆抗体 R2D6 和 A2B5 的反应所揭示的。这些发现表明 INS-1 细胞保持稳定并保留高度分化，这应该使它们成为研究 β 细胞功能各个方面的合适模型。
New insulin-secreting cell lines (INS-1 and INS-2) were established from cells isolated from an x-ray-induced rat transplantable insulinoma. The continuous growth of these cells was found to be dependent on the reducing agent 2-mercaptoethanol. Removal of this thiol compound caused a 15-fold drop in total cellular glutathione levels. These cells proliferated slowly (population doubling time about 100 h) and, in general, showed morphological characteristics typical of native beta-cells. Most cells stained positive for insulin and did not react with antibodies against the other islet hormones. The content of immunoreactive insulin was about 8-mu-g/10(6) cells, corresponding to 20% of the native beta-cell content. These cells synthesized both proinsulin I and II and displayed conversion rates of the two precursor hormones similar to those observed in rat islets. However, glucose failed to stimulate the rate of proinsulin biosynthesis. In static incubations, glucose stimulated insulin secretion from floating cell clusters or from attached cells. Under perifusion conditions, 10 mM but not 1 mM glucose enhanced secretion 2.2-fold. In the presence of forskolin and 3-isobutyl-1-methylxanthine, increase of glucose concentration from 2.8-20 mM caused a 4-fold enhancement of the rate of secretion. Glucose also depolarized INS-1 cells and raised the concentration of cytosolic Ca2+. This suggests that glucose is still capable of eliciting part of the ionic events at the plasma membrane, which leads to insulin secretion. The structural and functional characteristics of INS-1 cells remained unchanged over a period of 2 yr (about 80 passages). Although INS-2 cells have not been fully characterized, their insulin content was similar to that of INS-1 cells and they also remain partially sensitive to glucose as a secretagogue. INS-1 cells retain beta-cell surface antigens, as revealed by reactivity with the antiganglioside monoclonal antibodies R2D6 and A2B5.These findings indicate that INS-1 cells have remained stable and retain a high degree of differentiation which should make them a suitable model for studying various aspects of beta-cell function.