Engineering vascularized tissue in vivo using postnatal progenitor cells

使用出生后祖细胞改造体内血管化组织

• 批准号: 8510643
• 负责人: Juan M Melero-Martin
• 金额: $ 22.71万
• 依托单位: BOSTON CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-08-15 至 2014-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8510643
• 关键词: Acceleration; Address; Adipocytes; Adipose tissue; Adopted; Advanced Development; Affect; Age; Animals; Appearance; Area; Award; Biochemical; Bioluminescence; Blood; Blood Vessels; Bone Marrow; Boston; Burn injury; Carcinoma; Cell Differentiation process; Cells; Clinical; Collagen; Confocal Microscopy; Congenital Abnormality; Data; Development; Diabetes Mellitus; Endocrinology; Engineering; Excision; Fatty acid glycerol esters; Fibrin; Filler; Generations; Goals; Hour; Human; Image; Immunofluorescence Immunologic; Implant; In Situ; In Vitro; Individual; Injury; Intramuscular; Knowledge; Label; Letters; Location; Luciferases; Mastectomy; Mesenchymal Stem Cells; Mus; Muscle Cells; Natural regeneration; Organ; Outcome; Patients; Pediatric Hospitals; Phase; Phenotype; Plastics; Population; Process; Publishing; Reconstructive Surgical Procedures; Recruitment Activity; Regenerative Medicine; Research; Site; Specificity; Staining method; Stains; Stem cells; Stimulus; Strawberry nevus; System; Techniques; Technology; Testing; Therapeutic; Time; Tissue Engineering; Tissues; Translating; Undifferentiated; Vascularization; base; blood perfusion; design; experience; implantation; improved; in vivo; in vivo Model; interest; lipid biosynthesis; matrigel; postnatal; research study; scale up; subcutaneous; success; tissue regeneration; tissue repair; tumor

The goal is to develop a cell-based technology that will allow engineering vascularized tissues in vivo by using postnatal progenitor cells that can be obtained from patients by non-invasive means. In particular, this proposal will focus on the combined use of bone marrow-derived mesenchymal progenitor cells (MPCs) and human blood-derived endothelial progenitor cells (EPCs) to form adipose tissue in vivo. The concept to test is whether implants containing highly purified and defined MPCs and EPCs will result in specialized vascularized tissues upon implantation in vivo. We hypothesize that for this to happen, 1) both MPCs and EPCs need first to create a vascular network that will allow blood perfusion through the implants; and 2) this vascularization will promote MPCs that have not assembled into the vascular network to undergo appropriate tissue development by adopting the phenotype ofthe surrounding tissue at the site of implantation. In this proposal, we will determine key parameters to accelerate the vasculogenic process to a time frame of 24-48 hours using the in vivo model we have established with human EPCs and MPCs. Next, we will determine whether the adipocytes found in the implants are human in origin or whether they are recruited into the implants from the host as part of an endogenous tissue repair process once a functional vascular network is provided. With this information, we will implement strategies to enhance the process of in vivo differentiation with the aim of having fully functional tissues. Finally, we will test this cell-based technology by evaluating whether we can create long-lasting adipose tissue pads that present low volume reduction over time. The expenmental approaches to address these aims include: 1) in vivo imaging of blood perfusion in the implants using luciferase-based bioluminescence 2) GFP labeling of implanted cells to track their specific contribution to the resulting tissues; 3) confocal microscopy to evaluate implants through immunofluorescence staining. We envision this two-cell, two-step system as an enabling technology that can be applied to many different tissues wherein functional blood vessels are essential.

目标是开发一种基于细胞的技术，通过以下方式在体内改造血管化组织： 使用可以通过非侵入性方式从患者身上获得的产后祖细胞。特别是，这 该提案将重点关注骨髓间充质祖细胞（MPC）和 人类血液来源的内皮祖细胞（EPC）在体内形成脂肪组织。要测试的概念 是含有高度纯化和明确的 MPC 和 EPC 的植入物是否会产生专门的 植入体内后的血管化组织。我们假设要发生这种情况，1）MPC 和 EPC首先需要创建一个血管网络，允许血液通过植入物灌注； 2）这个 血管化会促进尚未组装成血管网络的 MPC 进行适当的 通过采用植入部位周围组织的表型来进行组织发育。在这个 根据提案，我们将确定关键参数，将血管生成过程加速到 24-48 的时间范围 使用我们用人类 EPC 和 MPC 建立的体内模型。接下来，我们将确定 植入物中发现的脂肪细胞是否源自人类，或者是否被招募到 一旦功能性血管网络恢复，来自宿主的植入物作为内源性组织修复过程的一部分 假如。有了这些信息，我们将实施增强体内分化过程的策略 目的是拥有功能齐全的组织。最后，我们将通过评估来测试这种基于细胞的技术 我们是否可以制造出持久的脂肪组织垫，随着时间的推移，其体积减少程度较低。这 实现这些目标的实验方法包括：1）植入物中血液灌注的体内成像 使用基于荧光素酶的生物发光 2) 植入细胞的 GFP 标记以追踪其具体贡献 至所得组织； 3) 共聚焦显微镜通过免疫荧光染色评估植入物。 我们设想这种两单元、两步系统作为一种使能技术，可以应用于许多不同的领域 其中功能性血管是必不可少的组织。