Leavens Karla F, Osorio-Quintero Catherine, Yeuteuh Elisabeth Ashlyne, Perez-Profeta Francesca T, Dattoli Anna Ada, Cardenas-Diaz Fabian L, French Deborah L, Gadue Paul
Department of Pediatrics, Perelman School of Medicine, University of Pennsylvania and Division of Endocrinology and Diabetes, The Children's Hospital of Philadelphia, Philadelphia, PA, USA.
Center for Cellular and Molecular Therapeutics, The Children's Hospital of Philadelphia, Philadelphia, PA, USA; Department of Pathology and Laboratory Medicine, The Children's Hospital of Philadelphia, Philadelphia, PA, USA.
Stem Cell Res. 2024 Dec;81:103559. doi: 10.1016/j.scr.2024.103559. Epub 2024 Sep 11.
Over the past decade, the use of human stem cell-derived β cells (SC-β cells) to model pancreatic β cell development, function and disease has become increasingly common. Though protocols are rapidly improving, current directed differentiation strategies do not yield a pure population of insulin-positive SC-β cells in vitro. Therefore, it is experimentally advantageous to have reporter lines that allow for live sorting of insulin-positive populations. To aid in these studies, we have knocked mNeonGreen fluorescent protein into the endogenous insulin locus of the commonly used H1 (WA01) human embryonic stem cell line.
在过去十年中,利用人干细胞衍生的β细胞(SC-β细胞)来模拟胰腺β细胞的发育、功能和疾病变得越来越普遍。尽管方案正在迅速改进,但目前的定向分化策略在体外无法产生纯的胰岛素阳性SC-β细胞群体。因此,拥有能够对胰岛素阳性群体进行活细胞分选的报告细胞系在实验上具有优势。为了辅助这些研究,我们已将mNeonGreen荧光蛋白敲入常用的H1(WA01)人胚胎干细胞系的内源性胰岛素基因座中。