3D heterogeneous islet organoid generation from human embryonic stem cells using a novel engineered hydrogel platform
3D heterogeneous islet organoid generation from human embryonic stem cells using a novel engineered hydrogel platform
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DOI：
10.1016/j.biomaterials.2018.05.031
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发表时间：
2018-09-01
期刊：
影响因子：
14
通讯作者：
Banerjee, Ipsita
中科院分区：
文献类型：
--
作者：
Candiello, Joseph;Grandhi, Taraka Sai Pavan;Banerjee, Ipsita
类器官表现出自发的器官特异性组织、功能和多细胞复杂性，本质上是特定体内器官系统的体外繁殖。最近的工作已经证明人类多能干细胞（hPSC）作为组织特异性类器官工程的可行再生细胞来源。这对于工程化胰岛类器官特别相关，因为最近在从人类多能干细胞产生功能性β样细胞方面取得了进展。在这项研究中，我们报告了再生胰岛类器官的精确大小和细胞异质性的具体工程，使用一种新的水凝胶系统，Amikagel。Amikagel促进人胚胎干细胞衍生的胰腺祖细胞（hESC-PP）受控和自发聚集成坚固的均质球状体。该平台进一步允许精细控制多个细胞群体的整合以产生异质球状体，这是复杂的类器官工程的必要条件。Amikagel诱导的hESC-PP球状体形成增强了胰岛特异性Pdx-1和NKX6.1基因和蛋白的表达，同时也增加了定向群体的百分比。进一步诱导hESC-PP球状体朝向成熟的β-样细胞，所述成熟的β-样细胞表现出增加的β-细胞特异性INS 1基因和C-肽蛋白表达，沿着功能性胰岛素产生以响应体外葡萄糖刺激。hESC-PP与生物学相关支持内皮细胞的进一步整合导致多细胞类器官，其表现出向胰岛特异性INS 1基因和C肽蛋白表达的自发成熟，以及显著发育的细胞外基质支持系统沿着。这些发现为胰岛类器官工程建立了理想的Amikagel促进平台。(C)2018爱思唯尔有限公司版权所有。
Organoids, which exhibit spontaneous organ specific organization, function, and multi-cellular complexity, are in essence the in vitro reproduction of specific in vivo organ systems. Recent work has demonstrated human pluripotent stem cells (hPSCs) as a viable regenerative cell source for tissue-specific organoid engineering. This is especially relevant for engineering islet organoids, due to the recent advances in generating functional beta-like cells from human pluripotent stem cells. In this study, we report specific engineering of regenerative islet organoids of precise size and cellular heterogeneity, using a novel hydrogel system, Amikagel. Amikagel facilitated controlled and spontaneous aggregation of human embryonic stem cell derived pancreatic progenitor cells (hESC-PP) into robust homogeneous spheroids. This platform further allowed fine control over the integration of multiple cell populations to produce heterogeneous spheroids, which is a necessity for complex organoid engineering. Amikagel induced hESC-PP spheroid formation enhanced pancreatic islet-specific Pdx-1 and NKX6.1 gene and protein expression, while also increasing the percentage of committed population. hESC-PP spheroids were further induced towards mature beta-like cells which demonstrated increased Beta-cell specific INS1 gene and C-peptide protein expression along with functional insulin production in response to in vitro glucose challenge. Further integration of hESC-PP with biologically relevant supporting endothelial cells resulted in multicellular organoids which demonstrated spontaneous maturation towards islet-specific INS1 gene and C-peptide protein expression along with a significantly developed extracellular matrix support system. These findings establish Amikagel -facilitated platform ideal for islet organoid engineering. (C) 2018 Elsevier Ltd. All rights reserved.