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Sintov, Elad

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Sintov

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Elad

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Sintov, Elad

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    Charting Cellular Identity During Human in Vitro β-Cell Differentiation
    (Springer Science and Business Media LLC, 2019-05) Veres, Adrian; Faust, Aubrey; Bushnell, Henry; Engquist, Elise; Kenty, Jennifer; Harb, George; Poh, Yeh-Chuin; Sintov, Elad; Gürtler, Mads; Pagliuca, Felicia; Peterson, Quinn; Melton, Douglas
    In vitro differentiation of human stem cells can produce pancreatic β-cells; the loss of this insulin-secreting cell type underlies type 1 diabetes. Here, as a step towards understanding this differentiation process, we report the transcriptional profiling of more than 100,000 human cells undergoing in vitro β-cell differentiation, and describe the cells that emerged. We resolve populations that correspond to β-cells, α-like poly-hormonal cells, non-endocrine cells that resemble pancreatic exocrine cells and a previously unreported population that resembles enterochromaffin cells. We show that endocrine cells maintain their identity in culture in the absence of exogenous growth factors, and that changes in gene expression associated with in vivo β-cell maturation are recapitulated in vitro. We implement a scalable re-aggregation technique to deplete non-endocrine cells and identify CD49a (also known as ITGA1) as a surface marker of the β-cell population, which allows magnetic sorting to a purity of 80%. Finally, we use a high-resolution sequencing time course to characterize gene-expression dynamics during the induction of human pancreatic endocrine cells, from which we develop a lineage model of in vitro β-cell differentiation. This study provides a perspective on human stem-cell differentiation, and will guide future endeavours that focus on the differentiation of pancreatic islet cells, and their applications in regenerative medicine.