Instrument-Free Single-Cell Resolution of Transcriptome in Mouse Pancreatic Islet Cells

Type 1 diabetes is characterized by an almost complete depletion of insulin-producing beta cells. Thus, regeneration of beta cells through beta-cell neogenesis represents a major therapeutic strategy to cure the disease. We previously reported a mouse model of severe insulin resistance that leads to high insulin secretion and massive beta-cell neogenesis. To understand the origins of the newly formed beta cells and the genic programs involved in high insulin secretion, we aimed at defining the pancreatic islet cells transcriptome at high resolution cell by cell. Here, we tested the compatibility of the novel instrument-free 3’ single-cell RNA-sequencing benchtop kit Asteria™, recently released by Scipio bioscience (Paris, France), on pancreatic islet cells isolated from adult mice, which underwent cell dissociation by trypsin treatment. In the Asteria™ procedure, differently from the microfluidic-based single-cell one, coupling of solid polymer-barcoded beads and cells occurs in homogenous phase in a reversible hydrogel named RevGel™. Such conditions are claimed by the vendor to be causing less mechanical stress to cells for an expected more preserved transcriptome. Successive steps were classic molecular biology and bioinformatic analysis using Scipio bioscience’s data analysis cloud-based software Cytonaut™. We profiled the transcriptome of 938 single cells from healthy mice and identified a total of 12 cell subtypes. SingleR annotation based on reference data revealed all the major cell types commonly described in the literature, including a well-represented beta cell population and some rare cell types such as ductal and stellate cells. We demonstrate 3’ single-cell RNA-sequencing using Scipio bioscience’s kit Asteria™ as an effective strategy to dissect the heterogeneity and the complexity of mouse pancreatic islets and identify the gene expression signatures of each cell. Disclosure D.G.Dimitrova: None. A.Liboz: None. S.Lipin: None. C.Beaupere: None. N.Roblot: None. J.Komatsu: None. G.Guillemain: None. B.Blondeau: None.