Cited by 552Pancreatic function and diabetesDiabetes and associated disordersPluripotent Stem Cells Research

Charting cellular identity during human in vitro β-cell differentiation

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Veres, Adrian, Faust, Aubrey L., Bushnell, Henry L., Engquist, Elise N., Kenty, Jennifer Hyoje-Ryu, Harb, George, Poh, Yeh-Chuin, Sintov, Elad +4 more·2019·Nature

Abstract

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.

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Crossref · 10.1038/s41586-019-1168-5

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  • SC-EC cells were identified, are absent from primary human islets, and resemble an endocrine cell type normally found in the intestines.

    “One of the most interesting findings from these scRNA-seq studies was the identification of SC-EC cells, which are absent from primary human islets and resemble an endocrine cell type normally found in the intestines.”

    From Developments in stem cell-derived islet replacement therapy for treating type 1 diabetes (2023) · cites this paper as [47]

    The SC-islets and SC-EC cells sections identify SC-EC cells and describe their resemblance to gut enterochromaffin cells. The Discussion says enterochromaffin cells have not previously been observed in studies of human islets, but also notes rare evidence suggesting resident pancreatic enterochromaffin cells; it does not establish that they are categorically absent from primary human islets. Page numbers are unavailable in our copy. (LLM, conf=0.86)

    Experimental — accuracy not yet measured
  • Current protocols also produce a significant percentage of stem cell-derived enterochromaffin cells (SC-ECs), which resemble a type of intestinal endocrine.

    “Current protocols also produce a significant percentage of stem cell-derived enterochromaffin cells (SC-ECs), which resemble a type of intestinal endocrine.47”

    From Developments in stem cell-derived islet replacement therapy for treating type 1 diabetes (2023) · cites this paper as [47]

    The ‘SC-islets contain four major cell types’ section identifies SC-EC cells as a major population in both modified protocols and says they most resemble enterochromaffin cells. The ‘SC-EC cells’ section suggests their presence in other β-cell protocols, but does not establish that those protocols produce a significant percentage of them. (LLM, conf=0.82)

    Experimental — accuracy not yet measured
  • SC-EC cells are characterized by the expression of enterochromaffin gene markers, such as TPH1 and LMX1A, as well as the secretion of serotonin rather than insulin.

    “These cells are characterized by the expression of enterochromaffin gene markers, such as TPH1 and LMX1A, as well as the secretion of serotonin rather than insulin.”

    From Developments in stem cell-derived islet replacement therapy for treating type 1 diabetes (2023) · cites this paper as [47]

    The “SC-EC cells” section reports TPH1 and LMX1A expression, serotonin content, and serotonin release from SC-islets; Extended Data Fig. 5 describes SC-EC cells as secreting serotonin. But the paper also reports low levels of insulin in SC-EC cells and does not establish that these cells secrete serotonin instead of insulin. (LLM, conf=0.86)

    Experimental — accuracy not yet measured
  • Hepatic, mesenchyme, and pancreatic exocrine cells are possible off-targets reported from SC-islet differentiations.

    “Hepatic, mesenchyme, and pancreatic exocrine are also possible off-targets reported from SC-islet differentiations47,51,75”

    From Developments in stem cell-derived islet replacement therapy for treating type 1 diabetes (2023) · cites this paper as [47]

    The “Fates of non-endocrine cells” section (page unavailable) reports cells differentiating toward pancreatic exocrine fates and populations expressing mesenchymal markers during SC-islet differentiation. It does not report hepatic off-target cells, so this source supports only part of the list. (LLM, conf=0.88)

    Experimental — accuracy not yet measured
  • In subsequent stages, high expression of PDX1 in combination with expression of NKX6-1 and PTF1A was predictive of endocrine induction potential.

    “In subsequent stages, high expression of PDX1 in combination with expression of NKX6-1 and PTF1A was predictive of endocrine induction potential.”

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Cites

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Authors
Veres, A., Faust, A. L., Bushnell, H. L., Engquist, E. N., Kenty, J. H.-R., Harb, G., Poh, Y.-C., Sintov, E., Gürtler, M., Pagliuca, F. W., Peterson, Q. P., & Melton, D. A.
Affiliations
  • Harvard University (US)
  • Semma Therapeutics (United States) (US)
  • Mayo Clinic (US)
  • Howard Hughes Medical Institute (US)
Corresponding author
Melton, Douglas A.
Venue
Nature
Published
2019-05-08
Volume
569
Issue
7756
Pages
368-373
ISSN
  • 0028-0836 (ISSN-L)
  • 1476-4687
OpenAlex
W2944658035
Cited by
552
References
9
Citations by year
2026: 72 · 2025: 68 · 2024: 81 · 2023: 81 · 2022: 75 · 2021: 86 · 2020: 61 · 2019: 27
FWCI
58.33
Type
journal article
Language
en
Open access
Locations
  • Nature (publishedVersion)
  • PubMed (publishedVersion)
  • PubMed Central (acceptedVersion)
  • Digital Access to Scholarship at Harvard (DASH) (Harvard University) (submittedVersion, cc-by)
  • RePEc: Research Papers in Economics (submittedVersion)
Funders
  • Howard Hughes Medical Institute
  • National Institute of General Medical Sciences (T32GM007226)
  • National Institute of Diabetes and Digestive and Kidney Diseases (UC4DK104159, UC4DK104165)
Topics
  • Health Sciences › Medicine › Surgery › Pancreatic function and diabetes
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Diabetes and associated disorders
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Pluripotent Stem Cells Research
Keywords
beta cell differentiation, human stem cells, pancreatic endocrine cells, gene expression profiling, enterochromaffin cells, pancreatic exocrine cells, beta-cell identity, beta cell maturation, magnetic cell separation, gene expression dynamics, type 1 diabetes, regenerative medicine
MeSH terms
RNA-Seq, Animals, Cell Differentiation, Cell Separation, Humans, Insulin, Male, Stem Cells, Biomarkers, Cell Lineage, Integrin alpha1, Insulin-Secreting Cells, Mice, Single-Cell Analysis

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