Embryonic stem cells differentiate into insulin-producing cells without selection of nestin-expressing cells
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We present a new strategy for the differentiation of embryonic stem (ES) cells into insulin-producing cells via a multi-step process without selection and induction of nestin-positive cells. During ES cell differentiation, transcript levels of genes characteristic of early and mature beta cells including Pdx1, Pax4, insulin and islet amyloid pancreatic peptide are up regulated. Islet-like clusters are characterized by expression of C-peptide, insulin and partially cytokeratin 19 as well as by ion channel activity similar to that found in embryonic beta cells. Cells of islet-like clusters show glucose-dependent insulin release at terminal stage. At an intermediate stage, nestin is partially co-expressed with C-peptide and cytokeratin 19, whereas islet-like clusters at the terminal stage are nestin-negative. We conclude that expression of nestin and cytokeratin 19 is a normal property of ES cells preceding differentiation into C-peptide/insulin-producing cells without any selection for nestin-positive phenotypes.
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Several methods of differentiation of ES cells to hepatic and insulin-producing cells have been described, but determining the precise identity of these cells is problematic due to a lack of suitable markers.
“While several methods of differentiation of ES cells to hepatic and insulin-producing cells have been described, determining the precise identity of these cells is problematic due to a lack of suitable markers [20-23].”
From A systematic screen for genes expressed in definitive endoderm by Serial Analysis of Gene Expression (SAGE) (2007) · cites this paper as [22]
The Discussion (page unavailable) describes several methods for producing insulin-positive cells from ES cells and cautions that some cells may absorb insulin from the medium rather than produce it, supporting a difficulty in identifying them. It does not discuss ES-cell differentiation into hepatic cells or attribute the identification difficulty to a lack of suitable markers. (LLM, conf=0.84)
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Self-citation
- Authors
- Blyszczuk, P., Asbrand, C., Rozzo, A., Kania, G., St-onge, L., Rupnik, M., & Wobus, A. M.
- Affiliations
- European Neuroscience Institute Göttingen (DE)
- Leibniz-Institut für Pflanzengenetik und Kulturpflanzenforschung (IPK) (DE)
- Corresponding author
- Wobus, Anna M.
- Venue
- The International Journal of Developmental Biology
- Publisher
- EHU Press
- Published
- 2004
- Volume
- 48
- Issue
- 10
- Pages
- 1095-1104
- ISSN
- 0214-6282 (ISSN-L)
- OpenAlex
- W2160827059
- Cited by
- 110
- References
- 57
- Citations by year
- 2022: 1 · 2021: 1 · 2018: 3 · 2017: 2 · 2016: 3 · 2015: 2 · 2014: 3 · 2013: 6 · 2012: 6
- FWCI
- 11.40
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Locations
- The International Journal of Developmental Biology (publishedVersion)
- PubMed (publishedVersion)
- Funders
- Deutsche Forschungsgemeinschaft
- Bundesministerium für Bildung und Forschung (01GN0106)
- Bundesministerium für Bildung, Wissenschaft und Forschung
- Topics
- Health Sciences › Medicine › Surgery › Pancreatic function and diabetes
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Pluripotent Stem Cells Research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Diabetes and associated disorders
- Keywords
- embryonic stem cells, insulin-producing cells, beta cell differentiation, nestin, cytokeratin 19, islet-like clusters, PDX1, Pax4, C-peptide, glucose-dependent insulin secretion, pancreatic development
- MeSH terms
- Animals, C-Peptide, Cell Differentiation, Electrophysiology, Embryo, Mammalian, Enzyme-Linked Immunosorbent Assay, Insulin, Intermediate Filament Proteins, Islets of Langerhans, Keratins, Male, Mice, Inbred BALB C, Microscopy, Fluorescence, Models, Biological, Nerve Tissue Proteins, Peptides, Phenotype, RNA, Messenger, Stem Cells, Streptozocin, Time Factors, Trans-Activators, Homeodomain Proteins, Patch-Clamp Techniques, Cell Culture Techniques, Reverse Transcriptase Polymerase Chain Reaction, Carbonic Anhydrase II, Mice, Nestin
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