Xin Yurong, Adler Christina, Kim Jinrang, Ding Yueming, Ni Min, Wei Yi, Macdonald Lynn, Okamoto Haruka
Regeneron Pharmaceuticals, Inc;
Regeneron Pharmaceuticals, Inc.
J Vis Exp. 2019 Jul 18(149). doi: 10.3791/59866.
Pancreatic islets comprise of endocrine cells with distinctive hormone expression patterns. The endocrine cells show functional differences in response to normal and pathological conditions. The goal of this protocol is to generate high-quality, large-scale transcriptome data of each endocrine cell type with the use of a droplet-based microfluidic single-cell RNA sequencing technology. Such data can be utilized to build the gene expression profile of each endocrine cell type in normal or specific conditions. The process requires careful handling, accurate measurement, and rigorous quality control. In this protocol, we describe detailed steps for human pancreatic islets dissociation, sequencing, and data analysis. The representative results of about 20,000 human single islet cells demonstrate the successful application of the protocol.
胰岛由具有独特激素表达模式的内分泌细胞组成。内分泌细胞在对正常和病理状况的反应中表现出功能差异。本方案的目标是利用基于微滴的微流控单细胞RNA测序技术生成每种内分泌细胞类型的高质量、大规模转录组数据。此类数据可用于构建正常或特定条件下每种内分泌细胞类型的基因表达谱。该过程需要谨慎操作、精确测量和严格的质量控制。在本方案中，我们描述了人胰岛解离、测序和数据分析的详细步骤。约20000个人单胰岛细胞的代表性结果证明了该方案的成功应用。
