Fletcher D J, Grogan W M, Barras E, Weir G C
Endocrinology. 1983 Nov;113(5):1791-8. doi: 10.1210/endo-113-5-1791.
Dispersed pancreatic islet cells were analyzed for their low forward angle light scatter using flow cytometry. The cells produced a distinct light scatter pattern which appeared to be a function of cell size and not cell granularity. RIA of hormone content of cells collected from different regions of the pattern revealed that glucagon- and somatostatin-containing cells were concentrated in regions of lower scatter intensity and that insulin-containing cells were more numerous in regions of higher intensity. Relative to the original cell suspension, these preparations were enriched 3-fold in glucagon and somatostatin content and 6-fold in insulin content. The function of intact islets, unsorted dispersed cells, and sorted dispersed cells was examined before and after 4 days of culture. Before culture, all of the dispersed cell populations had elevated basal secretion compared with intact islets and did not respond to stimulatory concentrations of glucose, arginine, or 3-isobutyl-1-methylxanthine. After culture for 4 days, basal secretion fell, and responsiveness returned. In both the A/D cell-enriched and the B cell-enriched cultured populations, the percentage of single cells was approximately 95%. The insulin release patterns from these populations were similar to those from intact islets and unsorted dispersed cells. Glucagon release from all of the dispersed cell populations far exceeded that from intact islets. This study suggests that the structural organization of islets influences A cell function, but a clear influence upon B cell function has not been demonstrated.
采用流式细胞术分析分散的胰岛细胞的低前向角光散射情况。这些细胞呈现出一种独特的光散射模式,该模式似乎是细胞大小而非细胞颗粒度的函数。对从该模式不同区域收集的细胞的激素含量进行放射免疫分析显示,含胰高血糖素和生长抑素的细胞集中在散射强度较低的区域,而含胰岛素的细胞在散射强度较高的区域数量更多。相对于原始细胞悬液,这些制剂中胰高血糖素和生长抑素的含量富集了3倍,胰岛素含量富集了6倍。在培养4天前后检测完整胰岛、未分选的分散细胞和分选的分散细胞的功能。培养前,与完整胰岛相比,所有分散细胞群体的基础分泌均升高,且对刺激浓度的葡萄糖、精氨酸或3 -异丁基-1 -甲基黄嘌呤无反应。培养4天后,基础分泌下降,反应性恢复。在富含A/D细胞和富含B细胞的培养群体中,单细胞的百分比约为95%。这些群体的胰岛素释放模式与完整胰岛和未分选的分散细胞相似。所有分散细胞群体的胰高血糖素释放远远超过完整胰岛。这项研究表明胰岛的结构组织影响A细胞功能,但尚未证明对B细胞功能有明显影响。