Engineering vascularized tissue in vivo using postnatal progenitor cells

使用出生后祖细胞在体内工程血管化组织

• 批准号: 7740989
• 负责人: Juan M Melero-Martin
• 金额: $ 9万
• 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 2011-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7740989
• 关键词: Address; Adipocytes; Adipose tissue; Adopted; Advanced Development; Affect; Age; Anastomosis - action; Animals; Appearance; Area; Biochemical; Biological Models; Bioluminescence; Bioreactors; Blood; Blood Vessels; Bone Marrow; Boston; Burn injury; Carcinoma; Cells; Clinical; Collagen; Confocal Microscopy; Congenital Abnormality; Data; Development; Diabetes Mellitus; Endocrinology; Engineering; Excision; Fatty acid glycerol esters; Fibrin; Figs - dietary; Filler; Fluorescence; Goals; Hemangioma; Hour; Human; Human Ubiquitin; Image; Immunodeficient Mouse; Immunofluorescence Immunologic; Implant; In Situ; In Vitro; Individual; Injury; Intramuscular; Knowledge; Label; Letters; Location; Luciferases; Mastectomy; Mesenchymal Stem Cells; Modeling; Mus; Muscle Cells; Natural regeneration; Organ; Outcome; Patients; Pediatric Hospitals; Perfusion; Phase; Phenotype; Plastics; Population; Principal Investigator; Process; Proteins; Publishing; Reconstructive Surgical Procedures; Recruitment Activity; Regenerative Medicine; Research; Retroviral Vector; SCID Mice; Site; Source; Specificity; Staining method; Stains; Stem cells; System; Techniques; Technology; Testing; Therapeutic; Time; Tissue Engineering; Tissues; Translating; Ubiquitin C; Umbilical Cord Blood; Undifferentiated; Vascularization; Work; Wound Healing; angiogenesis; base; blood perfusion; design; experience; implantation; improved; in vivo; in vivo Model; interest; lipid biosynthesis; matrigel; perilipin A; postnatal; promoter; regenerative; research study; scale up; subcutaneous; success; tissue regeneration; tumor; vasculogenesis

DESCRIPTION (provided by applicant): My 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. I 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 of the surrounding tissue at the site of implantation. In this proposal, I 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, I 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, I will implement strategies to enhance the process of in vivo differentiation with the aim of having fully functional tissues. Finally, I 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 experimental 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. I 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. RELEVANCE: This is an enabling technology that can be applied to regeneration of tissues/organs wherein functional blood vessels are essential. This proposal will focus on formation of adipose tissue, but it will be applicable to many aspects of regenerative medicine, including tissue-engineering and in situ therapeutic vascularization.

描述（由申请人提供）：我的目标是开发一种基于细胞的技术，该技术将允许通过使用可以通过非侵入性手段从患者获得的出生后祖细胞在体内工程化血管化组织。特别地，该提议将集中于骨髓来源的间充质祖细胞（MPC）和人血液来源的内皮祖细胞（EPC）的组合使用以在体内形成脂肪组织。测试的概念是含有高度纯化和确定的MPC和EPC的植入物是否会在体内植入后产生专门的血管化组织。我假设，要实现这一点，1）MPC和EPCs都需要首先创建一个血管网络，允许血液通过植入物灌注; 2）这种血管化将促进尚未组装成血管网络的MPC通过采用植入部位周围组织的表型进行适当的组织发育。在这个建议中，我将确定关键参数，以加速血管生成过程的时间框架为24-48小时，使用体内模型，我们已经建立了与人类EPCs和MPCs。接下来，我将确定在植入物中发现的脂肪细胞是否是人类起源的，或者它们是否是从宿主招募到植入物中的，作为一旦功能性血管网络被破坏后内源性组织修复过程的一部分。 提供了有了这些信息，我将实施策略，以增强体内分化的过程，目的是拥有功能齐全的组织。最后，我将测试这种基于细胞的技术，评估我们是否可以创造出持久的脂肪组织垫，随着时间的推移，脂肪组织垫的体积会减少。解决这些目标的实验方法包括：1）使用基于谷胱甘肽转移酶的生物发光对植入物中的血液灌注进行体内成像; 2）植入细胞的GFP标记以跟踪它们对所得组织的特定贡献; 3）共聚焦显微镜以通过免疫荧光染色来评估植入物。我设想这种两细胞，两步系统作为一种使能技术，可以应用于许多不同的组织，其中功能性血管是必不可少的。 相关性：这是一种可应用于组织/器官再生的技术，其中功能性血管是必不可少的。这项建议将集中在脂肪组织的形成，但它将适用于再生医学的许多方面，包括组织工程和原位治疗血管化。