Clardy Susan M, Mohan James F, Vinegoni Claudio, Keliher Edmund J, Iwamoto Yoshiko, Benoist Christophe, Mathis Diane, Weissleder Ralph
Center for Systems Biology, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts.
Division of Immunology, Department of Microbiology and Immunobiology, Harvard Medical School, Boston, Massachusetts.
Sci Rep. 2015 Sep 2;5:13681. doi: 10.1038/srep13681.
The ability to isolate pure pancreatic ß-cells would greatly aid multiple areas of diabetes research. We developed a fluorescent exendin-4-like neopeptide conjugate for the rapid purification and isolation of functional mouse pancreatic β-cells. By targeting the glucagon-like peptide-1 receptor with the fluorescent conjugate, β-cells could be quickly isolated by flow cytometry and were >99% insulin positive. These studies were confirmed by immunostaining, microscopy and gene expression profiling on isolated cells. Gene expression profiling studies of cytofluorometrically sorted β-cells from 4 and 12 week old NOD mice provided new insights into the genetic programs at play of different stages of type-1 diabetes development. The described isolation method should have broad applicability to the β-cell field.
分离纯的胰腺β细胞的能力将极大地有助于糖尿病研究的多个领域。我们开发了一种荧光艾塞那肽-4样新肽偶联物，用于快速纯化和分离功能性小鼠胰腺β细胞。通过用荧光偶联物靶向胰高血糖素样肽-1受体，可通过流式细胞术快速分离β细胞，且胰岛素阳性率>99%。这些研究通过对分离细胞的免疫染色、显微镜检查和基因表达谱分析得到了证实。对4周龄和12周龄非肥胖糖尿病（NOD）小鼠经细胞荧光分选的β细胞进行的基因表达谱研究，为1型糖尿病发展不同阶段所涉及的遗传程序提供了新的见解。所描述的分离方法应在β细胞领域具有广泛的适用性。
