Dynamic double network hydrogel for generating pancreatic organoids from induced pluripotent stem cells

动态双网络水凝胶用于从诱导多能干细胞生成胰腺类器官

• 批准号: 10636859
• 负责人: Chien-Chi Lin
• 金额: $ 46.85万
• 依托单位: INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-01 至 2026-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10636859
• 关键词: 3-Dimensional; Acinar Cell; Adhesives; Adopted; Affect; Basement membrane; Basic Science; Biochemical; Cell Differentiation process; Cell Therapy; Cells; Chemicals; Chemistry; Complex; Development; Diels Alder reaction; Disease model; Duct (organ) structure; Ductal Epithelial Cell; Ectoderm; Elasticity; Electrons; Encapsulated; Endoderm; Engineering; Esters; Exhibits; Exocrine pancreas; Extracellular Matrix; Future; Gel; Gelatin; Generations; Germ Layers; Glycols; Goals; Human; Hydrogels; In Situ; In Vitro; Integrin alpha5beta1; Integrins; Knowledge; Ligands; Mechanics; Mediating; Mesoderm; Modeling; Organogenesis; Organoids; Pancreas; Pancreatic Diseases; Pancreatic duct; Patients; Peptides; Pilot Projects; Polymers; Property; Proteins; Reaction; Regenerative Medicine; Relaxation; Role; Source; Specific qualifier value; Stress; Surface; Technology; Testing; Tissues; Transcription Coactivator; Trees; Work; biophysical properties; design; differentiation protocol; drug testing; functional group; improved; induced pluripotent stem cell; innovation; islet stem cells; mechanical properties; novel; progenitor; stem cell biology; stem cell fate; stem cell niche; success; therapeutic evaluation; tissue regeneration; translational applications; two-dimensional; viscoelasticity

PROJECT SUMMARY Human induced pluripotent stem cells (hiPSCs) can be differentiated to cells in all three germ layers (ectoderm, mesoderm, and endoderm), providing an invaluable cell source for basic research and translational applications. While recent years have witnessed breakthroughs in lineage-specific differentiation of hiPSC, the effect of matrix stiffness, viscoelasticity, and integrin ligand presentation during multi-stage exocrine pancreatic organoid (exoPO) development remain largely unexplored. Furthermore, current three-dimensional (3D) matrices for hiPSC culture and differentiation do not provide sufficient controls over matrix biophysical properties (e.g., viscoelasticity, stiffness) and biochemical motifs (e.g., cell-adhesive ligands). Additionally, no prior work has employed dynamic xeno-free hydrogels to study the effect of matrix mechanics and cell- adhesive ligand presentation on the development of hiPSC-derived exoPO. We hypothesize that exoPO differentiation can be drastically improved by presenting the cells with fine-tuned 3D matrix properties during the developmental stages. To achieve this goal, we will develop a viscoelastic dynamic double network (DDN) hydrogel platform with unprecedented tunability in matrix mechanical properties and biochemical motifs. Specifically, we will control matrix stiffness by forming an elastic hydrogel network with inverse Electron Demand Diels-Alder (iEDDA) click reaction. We will tune matrix stress-relaxation through a set of linear polymers complexed by reversible boronate-diol bonding. Uniquely, the elastic iEDDA click hydrogel network will be engineered to exhibit tunable hydrolytic degradation. On the other hand, the viscoelastic network will allow conjugation of cell adhesive ligands to permit viscoelasticity mediated engagement of integrins. With this viscoelastic DDN hydrogel platform, we will define the impact of matrix viscoelasticity, stiffness, and integrin ligand presentation on multipotent pancreatic progenitor cell differentiation and exoPO formation. In Aim 1, we will study the role of matrix viscoelasticity on pancreatic progenitor differentiation. In Aim 2, we will describe the requirements of matrix stiffness during pancreatic progenitor differentiation. In Aim 3, we will identify the role of cell adhesive ligands on pancreatic ductal/acinar cell specification. In the long-term, this project will produce a dynamic hydrogel platform to advance the use of chemically-defined matrices as xeno-free artificial stem cell niches for organoid development and tissue regeneration applications.

项目摘要 人诱导多能干细胞（hiPSC）可以分化为所有三个胚层中的细胞 （外胚层，中胚层和内胚层），为基础研究和转化提供了宝贵的细胞来源。 应用.虽然近年来hiPSC的谱系特异性分化已经有了突破，但在hiPSC的分化中， 多阶段胰腺外分泌过程中基质硬度、粘弹性和整合素配体呈递的影响 类器官（exoPO）的发育仍然在很大程度上未被探索。此外，当前的三维（3D） 用于hiPSC培养和分化的基质不能提供对基质生物物理的充分控制 属性（例如，粘弹性，硬度）和生物化学基序（例如，细胞粘附配体）。此外，没有 先前的工作已经使用动态的无异种水凝胶来研究基质力学和细胞的影响， 粘附配体呈现对hiPSC衍生的exoPO的发展的影响。我们假设exoPO 通过在细胞分化过程中呈现具有微调的3D基质性质的细胞， 发展阶段。为了实现这一目标，我们将开发一种粘弹性动态双网络（DDN） 水凝胶平台在基质机械性能和生物化学基序方面具有前所未有的可调节性。 具体来说，我们将通过形成具有逆电子的弹性水凝胶网络来控制基质刚度。 要求Diels-Alder（iEDDA）点击反应。我们将通过一组线性的 通过可逆的硼酸酯-二醇键合络合的聚合物。独特的是，弹性iEDDA点击水凝胶网络 将被设计成表现出可调节的水解降解。另一方面，粘弹性网络将 允许细胞粘附配体的结合以允许整合素的粘弹性介导的接合。与此 粘弹性DDN水凝胶平台，我们将定义基质粘弹性，刚度和整合素 配体呈递对多能胰腺祖细胞分化和exoPO形成的影响。目标1： 将研究基质粘弹性对胰腺祖细胞分化的作用。在目标2中，我们将描述 胰腺祖细胞分化过程中基质硬度的要求。在目标3中，我们将确定 细胞粘附配体对胰腺导管/腺泡细胞特化的影响。从长远来看，该项目将产生一个 动态水凝胶平台，以促进化学定义的基质作为无异种人工干细胞的使用 用于类器官发育和组织再生应用的小生境。