Generation of Functional Human Pancreatic β Cells In Vitro
Open accessAbstract
The generation of insulin-producing pancreatic β 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 β cells. Here, we report a scalable differentiation protocol that can generate hundreds of millions of glucose-responsive β cells from hPSC in vitro. These stem-cell-derived β cells (SC-β) express markers found in mature β cells, flux Ca(2+) in response to glucose, package insulin into secretory granules, and secrete quantities of insulin comparable to adult β 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.
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Transplantation of SC-islets into mice improves their insulin secretion and glucose responsiveness, suggesting that SC-islet immaturity can be resolved.
“Transplantation of SC-islets into mice improves their insulin secretion and glucose responsiveness, suggesting that SC-islet immaturity can be resolved.”
From Developments in stem cell-derived islet replacement therapy for treating type 1 diabetes (2023) · cites this paper as [36]
The Results section, “Stem Cell-Derived β Cells Function in vivo After Transplantation” (page unavailable), reports human insulin secretion and a response to glucose within two weeks of transplantation. It does not show that transplantation improved either property relative to the cells before transplantation. The separate Results section on in vitro function reports that SC-β cells were already glucose-responsive, so the proposed resolution of their immaturity is not established here. (LLM, conf=0.87)
Experimental — accuracy not yet measuredThe largest SC-islet differentiation reported in the literature was performed in 500 mL magnetic spinner flasks.
“and the largest SC-islet differentiation that has been reported in literature was performed in 500 mL magnetic spinner flasks.36”
From Developments in stem cell-derived islet replacement therapy for treating type 1 diabetes (2023) · cites this paper as [36]
The Discussion (page unavailable) reports growing 300 million cells at a time in suspension in a single 500 mL flask. Experimental Procedures > Cell culture (page unavailable) describes 500 mL spinner flasks on a stir plate. The paper does not establish that this was the largest SC-islet differentiation reported in the literature, and it does not explicitly call the flasks magnetic. (LLM, conf=0.84)
Experimental — accuracy not yet measuredStem cells begin as proliferative undifferentiated cells that are exposed to a series of cycles of growth and differentiation factors that lead to seven defined stages.
“These stem cells begin as proliferative undifferentiated cells that are exposed to a series of cycles of growth and differentiation factors that lead to seven defined stages.”
From The future of type 1 diabetes therapy (2025) · cites this paper as [68]
The Experimental Procedures (Cell culture; page not provided) begin with undifferentiated human pluripotent stem cells and describe successive changes in differentiation media. The Results (Generation of Glucose-Sensing Insulin-Secreting β Cells In Vitro; page not provided) also describe sequential culture steps using growth factors and other signaling agents. Our copy does not establish the claimed count of seven defined stages. (LLM, conf=0.84)
Experimental — accuracy not yet measuredIn 2014-15, several groups published protocols for generating stem cell-derived β (SC-β) cells that secreted insulin in response to glucose stimulation.
“In 2014-15, several groups published protocols for generating stem cell-derived β (SC-β) cells that secreted insulin in response to glucose stimulation.36–38”
From Developments in stem cell-derived islet replacement therapy for treating type 1 diabetes (2023) · cites this paper as [36]
These SC-β cells have been shown to improve glycemic control in diabetic mice.
“These cells have been shown to improve glycemic control in diabetic mice, highlighting their potential utility in vivo.36,37,39–44”
From Developments in stem cell-derived islet replacement therapy for treating type 1 diabetes (2023) · cites this paper as [36]
SC-islet protocols performed completely in suspension culture benefit from the natural three-dimensional scaling of these systems.
“SC-islet protocols that are performed completely in suspension culture36 benefit from the natural three-dimensional scaling of these systems.”
From Developments in stem cell-derived islet replacement therapy for treating type 1 diabetes (2023) · cites this paper as [36]
Methods have been developed by which pluripotent stem cells can be differentiated into β-like cells by a series of steps in which differentiation factors are added.
“Methods have been developed (figure 2) by which pluripotent stem cells can be differentiated into βlike cells by a series of steps in which differentiation factors are added.68,69”
From The future of type 1 diabetes therapy (2025) · cites this paper as [68]
The first four stages produce proliferative cells; as the cells differentiate, they become less proliferative.
“The first four stages produce proliferative cells.As the cells differentiate, they become less proliferative.”
From The future of type 1 diabetes therapy (2025) · cites this paper as [68]
The Results section describes pancreatic progenitors differentiating into β-like cells, and Experimental Procedures > Cell culture lists the stages of the protocol (page numbers unavailable). Neither reports that cells in the first four stages are proliferative or that proliferation declines with differentiation; searches for “proliferative,” “proliferation,” and cell-cycle terms found no such comparison. (LLM, conf=0.74)
Experimental — accuracy not yet measuredPagliuca and colleagues kept human embryonic stem cells suspended in liquid culture at 70 rpm, added specific growth factors to encourage differentiation, and found that the resulting beta-like cells behaved similarly to mature beta cells.
“For instance, Pagliuca and colleagues kept human embryonic stem cells suspended in liquid culture at 70 rpm, added specific growth factors to encourage differentiation, and found that the resulting beta-like cells behaved similarly to mature beta cells220.”
From Towards organoid culture without Matrigel (2021) · cites this paper as [220]
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- Authors
- Pagliuca, F. W., Millman, J. R., Gürtler, M., Segel, M., Van Dervort, A., Ryu, J. H., Peterson, Q. P., Greiner, D., & Melton, D. A.
- Affiliations
- Harvard University (US)
- University of Massachusetts Chan Medical School (US)
- Corresponding author
- Melton, Douglas A.
- ORCID
- Venue
- Cell
- Publisher
- Elsevier BV
- Published
- 2014-10
- Volume
- 159
- Issue
- 2
- Pages
- 428-439
- ISSN
- 0092-8674 (ISSN-L)
- PMC
- PMC4617632
- OpenAlex
- W2148890861
- Cited by
- 2,025
- References
- 50
- Citations by year
- 2026: 104 · 2025: 147 · 2024: 152 · 2023: 159 · 2022: 183 · 2021: 196 · 2020: 185 · 2019: 168 · 2018: 175 · 2017: 181 · 2016: 200 · 2015: 157 · 2014: 16
- FWCI
- 183.24
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Licence
- Locations
- Cell (publishedVersion)
- PubMed (publishedVersion)
- Europe PMC (PubMed Central) (acceptedVersion)
- PubMed Central (acceptedVersion)
- Funders
- Howard Hughes Medical Institute
- National Institutes of Health (1uc4dk104218-01)
- Harvard Stem Cell Institute
- National Institute of Diabetes and Digestive and Kidney Diseases (UC4-DK-104218, U01DK089572, P30DK020593)
- 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
- pancreatic beta cells, human pluripotent stem cells, stem cell-derived beta cells, glucose-dependent insulin secretion, beta cell differentiation, insulin secretion, calcium flux, cell transplantation, diabetes mellitus, hyperglycemia, secretory granules
- MeSH terms
- Animals, Humans, Insulin, Islets of Langerhans, Cell Culture Techniques, Pluripotent Stem Cells, Insulin-Secreting Cells, Mice, Induced Pluripotent Stem Cells
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