An Autologous, Culture-Free, Adipose Cell-Based Tissue Engineered Vascular Graft

一种自体、无培养、基于脂肪细胞的组织工程血管移植物

• 批准号: 9015874
• 负责人: David Alan Vorp
• 金额: $ 19.18万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-04-15 至 2018-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9015874
• 关键词: Acute; Address; Adipocytes; Adipose tissue; Animals; Autologous; Biological; Bioreactors; Blood Vessels; Bypass; Caliber; Cardiovascular Diseases; Cardiovascular system; Cell Culture Techniques; Cell Fraction; Cells; Clinic; Clinical; Collagen; Culture Media; Data; Devices; Elastin; Endothelial Cells; Evaluation; Fatty acid glycerol esters; Gold; Harvest; Homeostasis; Hour; Human; Human Resources; Implant; In Vitro; Lead; Length; Length of Stay; Mechanics; Mesenchymal Stem Cells; Modeling; Outcome; Paired Comparison; Patients; Performance; Pericytes; Population; Preparation; Process; Property; Proteins; Protocols documentation; Rattus; Risk; Seeds; Smooth Muscle Myocytes; Source; Stem cells; Suction Lipectomy; Technology; Testing; Time; Tissue Engineering; Tissues; Translations; Tubular formation; Vacuum; Vascular Graft; Vascular Smooth Muscle; Work; base; cell transformation; cost; design; human subject; implantation; in vivo; innovation; new technology; novel; public health relevance; research clinical testing; scaffold; scale up; vascular tissue engineering

 DESCRIPTION: Small-diameter tissue engineered vascular grafts (TEVGs) that incorporate autologous cells may provide an immuno-compatible treatment solution for patients with cardiovascular disease. We and others have shown that mesenchymal stem cells can promote remodeling of a biodegradable scaffold into native-like tissue that includes important vascular matrix proteins, such as collagen and elastin, and cells which contribute to vascular homeostasis, such as vascular smooth muscle cells and endothelial cells. Most recently, we have created TEVGs in this manner using adipose-derived mesenchymal stem cells from human donors. This application will address three critical barriers to translation of our current TEVG technology to the clinic. First, in our current TEVG paradigm, adipose-derived stem cells (AD-MSCs) are culture-expanded before seeding into biodegradable scaffolds. This presents a concern for the cells transforming or becoming contaminated, and also could represent significant costs with respect to personnel and materials. In addition, the presence of animal-sourced culture media supplements is a regulatory hurdle for any biological technology. Second, while the dynamic culture of a seeded graft for 48 hours - also part of our current TEVG protocol - is rapid compared to alternate approaches, it carries the same costs and risks as stated above for cell culture. Finally, our current TEVG has been optimized as an aortic interposition graft in the rat. Creating "human-sized" TEVG constructs for clinical translation wil require "scale-up" of at least one important process: seeding of cells into the scaffold. To address these concerns and advance our TEVG technology closer toward clinical feasibility, we propose the following three Specific Aims: 1) Evaluate the potential of using cells directly harvested from the stromal vascular fraction (SVF) immediately following liposuction (i.e., without time in culture) for fabricating our TEVG; 2) Eliminate the need for in-vitro bioreactor culture of our seeded construct to yield a TEVG that is safely and rapidly fabricated and ready to implant; and 3) Develop a new bulk-seeding device with the ability to deliver cells at high efficiency into "human-sized" scaffolds. Upon completion of this study, we expect to have developed an innovative "bedside" TEVG fabrication strategy using an alternative human adipose-derived cell source and a novel bulk-seeding device for long tubular scaffolds, and requiring no in vitro culture period. This will provide the basis of a new, innovative paradigm in vascular tissue engineering by eliminating significant regulatory barriers to clinical translation.

 描述：结合自体细胞的小直径组织工程血管移植物（TEVG）可以为心血管疾病患者提供免疫相容的治疗解决方案。我们和其他人已经证明，间充质干细胞可以促进可生物降解的支架重塑为天然样组织，其中包括重要的血管基质蛋白（例如胶原蛋白和弹性蛋白）以及有助于血管稳态的细胞（例如血管平滑肌细胞和内皮细胞）。最近，我们使用来自人类捐赠者的脂肪间充质干细胞以这种方式创建了 TEVG。 该应用程序将解决将我们当前的 TEVG 技术转化为临床的三个关键障碍。首先，在我们当前的 TEVG 范例中，脂肪源性干细胞 (AD-MSC) 在接种到可生物降解的支架中之前进行培养扩增。这引起了对细胞转化或被污染的担忧，并且还可能代表人员和材料方面的巨大成本。此外，动物源性培养基补充剂的存在是任何生物技术的监管障碍。其次，虽然对接种的移植物进行 48 小时的动态培养（也是我们当前 TEVG 方案的一部分）与其他方法相比速度较快，但它具有与上述细胞培养相同的成本和风险。最后，我们目前的 TEVG 已被优化为大鼠的主动脉介入移植物。创建用于临床转化的“人体大小”TEVG 构建体将需要“放大”至少一个重要过程：将细胞接种到支架中。 为了解决这些问题并使我们的 TEVG 技术更接近临床可行性，我们提出以下三个具体目标： 1) 评估在吸脂后立即使用从基质血管部分 (SVF) 直接收获的细胞（即无需培养时间）来制造我们的 TEVG 的潜力； 2) 无需对我们的种子结构进行体外生物反应器培养，即可安全、快速地制造并准备植入的 TEVG； 3) 开发一种新的批量接种装置，能够高效地将细胞输送到“人体大小”的支架中。 这项研究完成后，我们期望开发出一种创新的“床边”TEVG 制造策略，使用替代的人类脂肪来源的细胞来源和用于长管状支架的新型批量接种装置，并且不需要体外培养期。通过消除临床转化的重大监管障碍，这将为血管组织工程的新的创新范式奠定基础。