Diabetes Center, University of California San Francisco, San Francisco, CA, USA.
Barbara Davis Center for Diabetes, University of Colorado, School of Medicine, Aurora, CO, USA.
Nat Cell Biol. 2019 Feb;21(2):263-274. doi: 10.1038/s41556-018-0271-4. Epub 2019 Feb 1.
Despite advances in the differentiation of insulin-producing cells from human embryonic stem cells, the generation of mature functional β cells in vitro has remained elusive. To accomplish this goal, we have developed cell culture conditions to closely mimic events occurring during pancreatic islet organogenesis and β cell maturation. In particular, we have focused on recapitulating endocrine cell clustering by isolating and reaggregating immature β-like cells to form islet-sized enriched β-clusters (eBCs). eBCs display physiological properties analogous to primary human β cells, including robust dynamic insulin secretion, increased calcium signalling in response to secretagogues, and improved mitochondrial energization. Notably, endocrine cell clustering induces metabolic maturation by driving mitochondrial oxidative respiration, a process central to stimulus-secretion coupling in mature β cells. eBCs display glucose-stimulated insulin secretion as early as three days after transplantation in mice. In summary, replicating aspects of endocrine cell clustering permits the generation of stem-cell-derived β cells that resemble their endogenous counterparts.
尽管在将人胚胎干细胞分化为产生胰岛素的细胞方面取得了进展，但在体外生成成熟的功能性β细胞仍然难以实现。为了实现这一目标，我们开发了细胞培养条件，以紧密模拟胰岛器官发生和β细胞成熟过程中发生的事件。特别是，我们专注于通过分离和重新聚集未成熟的β样细胞来形成胰岛大小的富集β细胞簇（eBC），从而重现内分泌细胞聚类。eBC 显示出类似于原代人β细胞的生理特性，包括强大的动态胰岛素分泌、对激动剂的钙信号增加以及线粒体供能的改善。值得注意的是，内分泌细胞聚类通过驱动成熟β细胞中刺激-分泌偶联的核心过程——线粒体氧化呼吸，诱导代谢成熟。eBC 在移植后三天即可进行葡萄糖刺激的胰岛素分泌。总之，复制内分泌细胞聚类的某些方面可以生成类似于内源性细胞的干细胞衍生β细胞。
