Proteomic analysis of decellularized pancreatic matrix identifies collagen V as a critical regulator for islet organogenesis from human pluripotent stem cells
Proteomic analysis of decellularized pancreatic matrix identifies collagen V as a critical regulator for islet organogenesis from human pluripotent stem cells
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DOI：
10.1016/j.biomaterials.2019.119673
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发表时间：
2020-03-01
期刊：
影响因子：
14
通讯作者：
Jin, Sha
中科院分区：
文献类型：
--
作者：
Bi, Huanjing;Ye, Kaiming;Jin, Sha
在胰腺组织工程中，从干细胞产生人类胰岛类器官一直具有挑战性，主要是由于对体外多细胞组织自组装所需的小生境的理解不足。在这项研究中，我们的目的是确定生物活性，化学定义的生态位，从天然的，生物材料的胰岛体外发育。我们采用先进的生物信息学分析方法研究了脱细胞大鼠胰腺细胞外基质（dpECM）水凝胶的蛋白质组学，并鉴定了dpECM水凝胶中稳定且丰富的V型胶原（ColV）。通过在基质包被基质中呈递ColV向人多能干细胞（iPSC）提供的小生境允许干细胞进展成由所有主要胰腺内分泌细胞类型（即α、β、δ和胰腺多肽细胞）组成的胰岛样类器官。在ColV小生境的存在下，所有关键胰腺转录因子和主要激素基因的基因表达在iPSC衍生的类器官中显著增加。最重要的是，含有ColV的微环境导致类器官的胰岛素和胰高血糖素激素两者的葡萄糖响应性分泌增强。该研究表明，ColV是增强iPSC胰岛自组装的关键调节因子，并且利用天然生物材料构建体外多细胞组织生产所必需的组织线索是可行的。
In pancreatic tissue engineering, generating human pancreatic islet organoids from stem cells has been challenging due mainly to a poor understanding of niches required for multicellular tissue self-assembly in vitro. In this study, we aimed to identify bioactive, chemically defined niches from natural, biological materials for islet development in vitro. We investigated the proteomics of decellularized rat pancreatic extracellular matrix (dpECM) hydrogel using advanced bioinformatics analysis, and identified that type V collagen (ColV) is constantly and abundantly present in dpECM hydrogel. Niches provided to human pluripotent stem cells (iPSCs) by presenting ColV in matrix coating substrates permitted stem cells progression into islet-like organoids that consist of all major pancreatic endocrine cell types, i.e. a, beta, delta, and pancreatic polypeptide cells. In the presence of ColV niches, gene expressions of all key pancreatic transcription factors and major hormone genes significantly increased in iPSC-derived organoids. Most importantly, ColV-containing microenvironment resulted in enhanced glucose responsive secretions of both insulin and glucagon hormone from organoids. The study demonstrates that ColV is a critical regulator that augments islet self-assembly from iPSCs, and it is feasible to utilize natural biomaterials to build tissue cues essential for multicellular tissue production in vitro.