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GSE70901
Homo sapiens
16 Downloadable Samples
Illumina HumanHT-12 V4.0 expression beadchip
Description
We recently reported the scalable in vitro production of functional stem cell-derived cells. Here we extend this approach to generate SC- cells from Type 1 diabetic patients (T1D), a cell type that is destroyed during disease progression and has not been possible to extensively study. These cells express cell markers, respond to glucose both in vitro and in vivo, prevent alloxan-induced diabetes in mice, and respond to anti-diabetic drugs. Furthermore, we use an in vitro disease model to demonstrate the cells respond to different forms of cell stress. Using these assays, we find no major differences in T1D SC- cells compared to SC- cells derived from non-diabetic patients (ND). These results show that T1D SC- cells can be used for the treatment of diabetes, drug screening, and the study of cell biology.
Publication Title
Generation of stem cell-derived β-cells from patients with type 1 diabetes.
Sample Metadata Fields
Specimen part
View Samples- Homo sapiens
- 2 Downloadable Samples
- Illumina HumanHT-12 V4.0 expression beadchip
Description
The generation of insulin-producing pancreatic cells from stem cells in vitro would provide an unprecedented cell source for drug discovery and cell transplantation therapy in diabetes. However, insulin-producing cells previously generated from human pluripotent stem cells (hPSC) lack many functional characteristics of bona fide cells. Here we report a scalable differentiation protocol that can generate hundreds of millions of glucose-responsive cells from hPSC in vitro. These stem cell derived cells (SC) express markers found in mature cells, flux Ca2+ in response to glucose, package insulin into secretory granules and secrete quantities of insulin comparable to adult cells in response to multiple sequential glucose challenges in vitro. Furthermore, these cells secrete human insulin into the serum of mice shortly after transplantation in a glucose-regulated manner, and transplantation of these cells ameliorates hyperglycemia in diabetic mice.
Publication Title
Generation of functional human pancreatic β cells in vitro.
Sample Metadata Fields
Specimen part, Cell line
View Samples- Mus musculus
- 684 Downloadable Samples
Illumina HiSeq 2500
Description
This scRNA-seq data is an integral part of a manuscript with the above title. Using computational methods, we were able to reconstruct a detailed branched trajectory reflecting pancreatic endocrine differentiation in the mouse embryo. Analysis of the transcriptional changes occuring during the differentiation suggested that epithelial-to-mesenchymal transition likely plays no role in this process, contrary to the prevailing dogma. Our findings were corroborated with high-resolution imaging of the developing pancreas, revealing how differentiating endocrine progenitors migrate in cohesion, forming bud-like islet precursors, or "peninsulas", and that spatiotemporal collinearity during differentiation leads to the typical core-mantle architecture of the mature, spherical islet. This work led to a complete overhaul of our understanding of how pancreatic islets are developed, laying the ground for the generation of entire islets in vitro as a potential novel source of islet transplantation. Overall design: Single-cell suspensions were prepared from pancreata of Neurogenin 3-eGFP mouse embryos sacrificed at different days of embryonic development. Single eGFP-positive cells were FACS-sorted into 96-well plates, and single-cell cDNA was prepared using the SMART-seq protocol. Single-cell sequencing libraries were generated using the Nextera XT DNA library preparation kit and sequenced on an Illumina HiSeq sequencer. Reads were aligned to the mouse reference genome build mm10 with TopHat, and single-cell gene expression profiles were computed using Cufflinks.
Publication Title
A Peninsular Structure Coordinates Asynchronous Differentiation with Morphogenesis to Generate Pancreatic Islets.
Sample Metadata Fields
Specimen part, Subject
View Samples- Homo sapiens
- 72 Downloadable Samples
- NextSeq 500
Description
In vitro differentiation of human stem cells can produce pancreatic beta cells, the insulin-secreting cell type whose loss underlies Type 1 Diabetes. As a step towards mastery of this process, we report on transcriptional profiling of >100,000 individual cells sampled during in vitro beta cell differentiation and describe the cells that emerge. We resolve populations corresponding to beta cells, alpha-like poly-hormonal cells, non-endocrine cells that resemble pancreatic exocrine cells and a previously unreported population resembling enterochromaffin cells. We show that the beta and alpha-like cells are stable for weeks in culture without exogenous growth factors and that gene expression changes associated with in vivo beta cell maturation are recapitulated in vitro. We demonstrate that stem-cell derived enterochromaffin cells can synthesize and secrete serotonin in vitro. To remove exocrine cells, we characterize a scalable re-aggregation technique that efficiently selects endocrine cells. Finally, we use a high-resolution sequencing time course to characterize gene expression dynamics during human pancreatic endocrine induction from which we develop a lineage model of in vitro beta cell differentiation. This study provides a deeper perspective on the current state of human stem cell differentiation and is a jumping-off point for future endeavors in in vitro differentiation of pancreatic islet cells and their application in regenerative medicine. Overall design: Single-cell mRNA sequencing of pluripotent stem cells differentiating in vitro towards pancreatic beta cells. The data & metadata match the initial submission of the manuscript, not the final version.
Publication Title
Charting cellular identity during human in vitro β-cell differentiation.
Sample Metadata Fields
Specimen part, Subject
View Samples