Andalusian Centre for Molecular Biology and Regenerative Medicine (CABIMER), CSIC, Pablo de Olavide University, Seville, Spain.
Department of Cell Biology, Genetics and Physiology, University of Malaga (UMA), The Institute of Biomedical Research in Malaga (IBIMA), Málaga, Spain.
In Vitro Cell Dev Biol Anim. 2021 Jun;57(6):587-597. doi: 10.1007/s11626-021-00580-8. Epub 2021 Jul 1.
Conventional methods for obtaining pancreatic β cells are based on simulating the embryonic development phase of endocrine cells via hierarchical differentiation of pluripotent stem cells (PSCs). Accordingly, we attempted to modify the protocols for obtaining insulin-secreting cells (ISCs) by sequential differentiation of a human embryonic stem cell (hESC), using the HS181 cell line. Furthermore, we hypothesize that actual pancreatic endocrine cells may arise from trans-differentiation of mature ductal cells after the embryonic developmental stage and throughout the rest of life. According to the hypothesis, ductal cells are trans-differentiated into endocrine and exocrine cells, undergoing a partial epithelial to mesenchymal transition (EMT). To address this issue, we developed two new protocols based on hESC differentiation to obtain ductal cells and then induce EMT in cells to obtain hormone-secreting islet-like cells (HSCs). The ductal (pre-EMT exocrine) cells were then induced to undergo partial EMT by treating with Wnt3a and activin A, in hypoxia. The cell derived from the latter method significantly expressed the main endocrine cell-specific markers and also β cells, in particular. These experiments not only support our hypothetical model but also offer a promising approach to develop new methods to compensate β cell depletion in patients with type 1 diabetes mellitus (T1DM). Although this protocol of generating islet-like cells from ductal cells has a potential to treat T1DM, this strategy may be exploited to optimize the function of these cells in an animal model and future clinical applications.
获得胰腺β细胞的传统方法基于通过多能干细胞（PSCs）的层次分化来模拟内分泌细胞的胚胎发育阶段。因此，我们试图通过顺序分化人胚胎干细胞（hESC）来修改获得胰岛素分泌细胞（ISCs）的方案，使用 HS181 细胞系。此外，我们假设实际的胰腺内分泌细胞可能是在胚胎发育阶段之后和整个生命过程中从成熟导管细胞的转分化而来。根据该假设，导管细胞转分化为内分泌和外分泌细胞，经历部分上皮间质转化（EMT）。为了解决这个问题，我们开发了两种基于 hESC 分化的新方案，以获得导管细胞，然后诱导 EMT 使细胞获得激素分泌胰岛样细胞（HSCs）。然后，通过在缺氧条件下用 Wnt3a 和激活素 A 处理，将这些导管细胞（前 EMT 外分泌细胞）诱导经历部分 EMT。来自后一种方法的细胞显著表达主要的内分泌细胞特异性标志物，特别是β细胞。这些实验不仅支持我们的假设模型，而且为开发新方法来补偿 1 型糖尿病（T1DM）患者的β细胞耗竭提供了有希望的途径。虽然这种从导管细胞生成胰岛样细胞的方案有可能治疗 T1DM，但该策略可用于优化这些细胞在动物模型和未来临床应用中的功能。
