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，该技术将通过使用可以通过非侵入性手段从患者那里获得的产后祖细胞在体内进行工程性血管化组织。特别是，该建议将集中于骨髓衍生的间充质祖细胞（MPC）和人血液来源的内皮祖细胞（EPC）在体内形成脂肪组织。要测试的概念是植入体内植入后植入高度纯化和定义的MPC和EPC是否会导致专门的血管组织。我假设要实现这一目标，1）MPC和EPC首先需要建立一个血管网络，该血管网络将使血液通过植入物进行血液灌注； 2）这种血管形成将促进尚未组装到血管网络中的MPC，以通过在植入部位采用周围组织的表型来进行适当的组织发育。在此提案中，我将使用使用人类EPC和MPC建立的体内模型来确定将血管生成过程加速到24-48小时的时间范围的关键参数。下一 假如。有了这些信息，我将实施策略来增强体内分化的过程，目的是具有功能齐全的组织。最后，我将通过评估我们是否可以创建持久的脂肪组织垫来测试这种基于细胞的技术，从而表现出较低的体积随时间的减少。解决这些目标的实验方法包括：1）使用基于荧光素酶的生物发光的植入物中血液灌注的体内成像2）植入细胞的GFP标记，以跟踪其对所得组织的特定贡献； 3）共聚焦显微镜通过免疫荧光染色评估植入物。我将这个两键两步的系统设想为一种启示技术，可以应用于功能性血管至关重要的许多不同组织。 相关性：这是一项可应用的技术，可以应用于功能血管至关重要的组织/器官的再生。该建议将集中于脂肪组织的形成，但它适用于再生医学的许多方面，包括组织工程和原位治疗性血管形成。