The First Affiliated Hospital, Jinan University, Guangzhou, China.
J Cell Biochem. 2013 Oct;114(10):2221-30. doi: 10.1002/jcb.24555.
Transplantation of functional insulin-producing cells (IPCs) provides a novel mode for insulin replacement, but is often accompanied by many undesirable side effects. Our previous studies suggested that IPCs could not mimic the physiological regulation of insulin secretion performed by pancreatic beta cells. To obtain a better method through which to acquire more similar IPCs, we compared the difference between IPCs of the GLP-1 group and IPCs of the non-GLP-1 group in the morphological features in cellular level and physiological function. The levels of insulin secretion were measured by ELISA. The insulin and glucagon-like peptide-1 (GLP-1) mRNA gene expression was determined by real-time quantitative PCR. The morphological features were detected by atomic force microscopy (AFM) and laser confocal scanning microscopy (LCSM). Intracellular Ca(2+) levels and Glucagon-like peptide-1 receptor (GLP-1R) levels were determined by flow cytometer (FCM). We found that IPCs of the GLP-1 group had bigger membrane particle size and average roughness (Ra ) than IPCs of the non-GLP-1 group but still smaller than normal human pancreatic beta cells. The physiology function of IPCs of the GLP-1 group were much closer to normal human pancreatic beta cells than IPCs of the non-GLP-1 group. GLP-1 could improve the similarity of IPCs from human adipose tissue-derived mesenchymal stem cells and pancreatic beta cells in cellular ultrastructure and function.
移植功能性胰岛素分泌细胞(IPCs)为胰岛素替代提供了一种新的模式,但常伴有许多不良的副作用。我们之前的研究表明,IPCs 不能模拟胰岛β细胞进行的胰岛素分泌的生理调节。为了获得更好的方法来获得更相似的 IPCs,我们比较了 GLP-1 组的 IPCs 和非 GLP-1 组的 IPCs 在细胞水平的形态特征和生理功能上的差异。通过 ELISA 测量胰岛素分泌水平。通过实时定量 PCR 测定胰岛素和胰高血糖素样肽-1(GLP-1)mRNA 基因表达。通过原子力显微镜(AFM)和激光共聚焦扫描显微镜(LCSM)检测形态特征。通过流式细胞仪(FCM)测定细胞内 Ca(2+)水平和胰高血糖素样肽-1 受体(GLP-1R)水平。我们发现 GLP-1 组的 IPCs 的膜颗粒大小和平均粗糙度(Ra)均大于非 GLP-1 组的 IPCs,但仍小于正常人胰腺β细胞。GLP-1 组的 IPCs 的生理功能比非 GLP-1 组的 IPCs 更接近正常人胰腺β细胞。GLP-1 可改善人脂肪组织来源的间充质干细胞和胰腺β细胞的 IPCs 在细胞超微结构和功能上的相似性。