Homo sapiens whole pancreatic islet mRNA during culture in alternate media

Two islet preparations (from different donors) were subjected to RNA-sequencing. Within each preparation, subsets of islets were treated with 5 or 8 mM glucose, or 8 mM glucose in a specialized, conditioned medium containing Wnt and R-spondin ligands with Rho and ROCK inhibitors (L-WRN+) for 48 h immediately after initial receipt and hand-selection. L-WRN+ promotes proliferation with undue de-differentiation. Polyadenylated mRNA was obtained from 1 mcg of human islet total RNA and 50 nt, single-end sequencing reads were obtained from an Illumina HiSeq 2500.

本研究采用两份来自不同供体的胰岛制备物开展RNA测序（RNA-sequencing）。每份制备物内的胰岛亚群均在初始接收并经手工分选后，即刻接受48小时的不同处理：分别使用5 mM葡萄糖、8 mM葡萄糖，或是在含有Wnt与R-spondin配体、同时添加Rho及ROCK抑制剂的专用条件培养基（L-WRN+）中添加8 mM葡萄糖。L-WRN+可促进胰岛细胞增殖，但会引发过度的去分化。从1微克（mcg）人胰岛总RNA中提取聚腺苷酸化mRNA，并通过Illumina HiSeq 2500测序平台生成50 nt的单端测序读段。