GSE121679
A peninsular structure coordinates asynchronous differentiation with morphogenesis to generate pancreatic islets [bulk vs buds]
This RNA-seq data is an integral part of a manuscript with the above title. To better characterize the buds appearing on the surface of the spheroidal clusters during the directed differentiation of HUES8 human embryonic stem cells to pancreatic endocrine cells, we performed bulk RNA-sequencing of the buds versus the bulks. Our results indicate that the buds indeed consist of pancreatic endocrine cells, illustrating our ability to recapitulate in vitro our in vivo observations of pancreatic islets forming via peninsular intermediates. See manuscript for more information. Overall design: Buds were detached and separated from the spheroids at stage 5 of the differentiation protocol by mechanical agitation and filtering. RNA-sequencing of buds versus bulks was performed on samples from 5 separate differentiation flasks, whereby bulks could be collected from all and buds from 3 of the differentiation flasks. Sequencing libraries were prepared using the PrepX RNA-Seq for Illumina Library Kit (Takara Bio USA) and sequenced on an Illumina NextSeq 500. Paired-end sequencing reads were aligned to the human genome version GRCh38 and quantified with RSEM using the STAR alignment algorithm with default settings.