期刊
FRONTIERS IN CELL AND DEVELOPMENTAL BIOLOGY
卷 10, 期 -, 页码 -出版社
FRONTIERS MEDIA SA
DOI: 10.3389/fcell.2022.967765
关键词
aggregate; beta cell; human induced pluripotent stem cell; insulin secretion; islet; microwell
资金
- Innovative Medicines Initiative 2 Joint Undertaking
- Union's Horizon 2020 research and innovation programme [115797, 945268]
- European Federation of Pharmaceutical Industries and Associations
- JDRF
- Leona M. and Harry B. Helmsley Charitable Trust
- European Union [1912-03555]
- EFSD/Boehringer Ingelheim European Research Programme on Multi-System Challenges in Diabetes [667191, 801505]
- Francophone Foundation for Diabetes Research - French Diabetes Federation
- Abbott
- Eli Lilly
- Merck Sharp and Dohme
- Novo Nordisk
- Fonds National de la Recherche Scientifique (FRS-FNRS)
- Brussels Capital Region-Innoviris (Bridge project DiaType)
- Walloon Region SPW-EER Win2Wal project BetaSource
- Fonds Erasme for Medical Research
- Fondation ULB
- Marie Curie Actions (MSCA) [801505] Funding Source: Marie Curie Actions (MSCA)
The study comprehensively characterized the functionality of iPSC-derived beta cells both in vitro and in vivo in humanized mice. Differentiation in microwells showed equal efficiency with rotating suspension, but with a higher success rate. In vivo transplanted beta cells demonstrated functional characteristics similar to human islets, highlighting the potential of generating islet-like organoids for diabetes research.
In vitro differentiation of human induced pluripotent stem cells (iPSCs) into beta cells represents an important cell source for diabetes research. Here, we fully characterized iPSC-derived beta cell function in vitro and in vivo in humanized mice. Using a 7-stage protocol, human iPSCs were differentiated into islet-like aggregates with a yield of insulin-positive beta cells comparable to that of human islets. The last three stages of differentiation were conducted with two different 3D culture systems, rotating suspension or static microwells. In the latter, homogeneously small-sized islet-like aggregates were obtained, while in rotating suspension size was heterogeneous and aggregates often clumped. In vitro function was assessed by glucose-stimulated insulin secretion, NAD(P)H and calcium fluctuations. Stage 7 aggregates slightly increased insulin release in response to glucose in vitro. Aggregates were transplanted under the kidney capsule of NOD-SCID mice to allow for further in vivo beta cell maturation. In transplanted mice, grafts showed glucose-responsiveness and maintained normoglycemia after streptozotocin injection. In situ kidney perfusion assays showed modulation of human insulin secretion in response to different secretagogues. In conclusion, iPSCs differentiated with equal efficiency into beta cells in microwells compared to rotating suspension, but the former had a higher experimental success rate. In vitro differentiation generated aggregates lacking fully mature beta cell function. In vivo, beta cells acquired the functional characteristics typical of human islets. With this technology an unlimited supply of islet-like organoids can be generated from human iPSCs that will be instrumental to study beta cell biology and dysfunction in diabetes.
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