Recellularization of Liver Bioscaffolds

肝脏生物支架的再细胞化

• 批准号: 7696149
• 负责人: Martin L Yarmush
• 金额: $ 41.17万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-05 至 2011-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7696149
• 关键词: Acute; Adrenal Cortex Hormones; Adult; Affect; Albumins; Ammonia; Animal Model; Apoptotic; Architecture; Biocompatible Materials; Blood Coagulation Factor; Cardiac Death; Cell Maturation; Cell Survival; Cell-Matrix Junction; Cells; Cessation of life; Chronic; Clinical; Coculture Techniques; Commit; Computer Assisted; Cues; Cytochrome P450; Cytoskeleton; Detection; Development; Drug Kinetics; Drug Metabolic Detoxication; Endothelial Cells; Engineering; Engraftment; Enhancers; Environment; Future; Gel; Goals; Graft Survival; Hepatic; Hepatic Mass; Hepatic Tissue; Hepatocyte; Hormones; Human; Image Analysis; In Vitro; Kidney; Lead; Liver; Liver Failure; Liver diseases; Lobe; Metabolic; Methods; Modeling; Mus; Organ; Organ Model; Organ Transplantation; Pancreas; Pathway interactions; Patients; Perfusion; Pharmacologic Substance; Positioning Attribute; Property; Protein C Inhibitor; Protocols documentation; Public Health; Rattus; Research; Serum; Solutions; Stem cells; System; Techniques; Testing; Time; Tissue Engineering; Tissues; Transplantation; Urea; Vascularization; Waiting Lists; Work; alpha 1-Antitrypsin; base; cell motility; cell type; cellular engineering; cytokine; embryonic stem cell; extracellular; graft function; hepatocyte engraftment; human embryonic stem cell; improved; in vivo; innovation; liver function; liver transplantation; novel; novel strategies; public health relevance; scaffold; stem cell differentiation; success

DESCRIPTION (provided by applicant): About thirty million people in the US undergo a liver disorder for different causes and about 27,000 deaths are registered annually in the US due to liver disease. At this time, the only definitive treatment of hepatic failure is orthotopic transplantation. However, there is a critical shortage of organs, with a deficit of ~3,000 livers per year. Similar numbers affect most organs & tissues, with the total organ waiting list currently at 100,000 requests and the number increasing by 5% every year. Given that only organs in pristine condition are transplantable, orthotopic transplantation will always remain a limited pool. A more elegant, long-term solution is using stem cells to develop tissue-engineered replacements. However, while many in vitro successes have been demonstrated, clinical success has been very limited due to low cell viability and functionality in the long term in vivo. The major gap is the lack of an ideal transplantable scaffold that has all the necessary microstructure and extracellular cues for cell attachment, differentiation, function and vascularization, which has so far proven difficult to manufacture in vitro. Our long-term goal is to engineer transplantable liver grafts for curing or treating relevant liver diseases. The objective of the proposed study is to develop functional and implantable liver grafts. The central hypothesis to be tested here is that the natural liver scaffold derived from discarded livers can be extensively repopulated, can provide an adequate maturation environment for stem cell derived liver cells, and that these grafts perform the essential hepatic functions in vivo. The rationale of the study is that while most research focuses on producing the ideal scaffold from the ground up using synthetic biomaterials, the native ECM is likely to contain the necessary architecture and environmental cues, hence presents a promising, little explored alternative approach for producing organ grafts which can vertically advance the field of tissue engineering. Engineering of functional liver grafts from stem-cell derived hepatocytes and liver's natural matrix is an innovative endeavor, as it has the potential to become a novel platform for hepatic tissue engineering. The work described here is expected to i) establish decellularized liver matrices as a viable scaffold for hepatic tissue engineering, ii) generate a liver ECM-based maturation protocol to generate hepatocyte-like cells, and iii) lead to a novel graft engineering approach to provide auxiliary hepatic support. While this work utilizes liver as the model organ, the results of this work will also have a positive impact by establishing the basis of future sophisticated organ engineering techniques that incorporate several different cell types and can be applied to other organs (pancreas, kidney, etc.), and may ultimately lead to development of entire organs in vitro. The ESC maturation protocol developed here is expected to be a significant contribution to the field of stem cell engineering. Hepatocyte culture in the decellularized matrix may also prove to be a new platform for pharmaceutical studies. PUBLIC HEALTH RELEVANCE: About thirty million people in the US undergo a liver disorder for different causes and about 27,000 deaths are registered annually in the US due to liver disease. At this time, the only definitive treatment of hepatic failure is orthotopic transplantation. However, there is a critical shortage of organs, with a deficit of ~3,000 livers per year. Similar numbers affect most organs & tissues, with the total organ waiting list currently at 100,000 requests and the number increasing by 5% every year. Given that only organs in pristine condition are transplantable, orthotopic transplantation will always remain a limited pool. The results of this study are expected to directly improve public health by the developing a novel approach for engineering auxiliary liver grafts using stem cell derived hepatocytes and native matrices from discarded livers, in order to treat patients with liver failure.

描述（由申请人提供）：美国约有 3000 万人因不同原因患有肝脏疾病，美国每年约有 27,000 人因肝脏疾病死亡。目前，肝衰竭唯一确定的治疗方法是原位移植。然而，器官严重短缺，每年短缺约 3,000 个肝脏。类似的数字影响到大多数器官和组织，目前器官等待名单总数为 100,000 个请求，并且该数字每年以 5% 的速度增长。鉴于只有处于原始状态的器官才可以移植，原位移植的数量始终有限。一个更优雅的长期解决方案是使用干细胞开发组织工程替代品。然而，虽然许多体外成功已被证明，但由于长期体内细胞活力和功能性较低，临床成功非常有限。主要差距是缺乏理想的可移植支架，该支架具有细胞附着、分化、功能和血管化所需的所有必要的微观结构和细胞外线索，迄今为止已证明很难在体外制造。我们的长期目标是设计可移植肝移植物来治愈或治疗相关的肝脏疾病。拟议研究的目的是开发功能性和可植入性肝移植物。这里要测试的中心假设是，源自废弃肝脏的天然肝脏支架可以广泛地重新填充，可以为干细胞衍生的肝细胞提供足够的成熟环境，并且这些移植物在体内执行基本的肝功能。该研究的基本原理是，虽然大多数研究重点是使用合成生物材料从头开始生产理想的支架，但天然 ECM 可能包含必要的结构和环境线索，因此提出了一种有前途的、很少探索的替代方法来生产器官移植物可以垂直推进组织工程领域。利用干细胞衍生的肝细胞和肝脏天然基质工程设计功能性肝移植物是一项创新工作，因为它有潜力成为肝组织工程的新平台。这里描述的工作预计将 i) 建立脱细胞肝脏基质作为肝组织工程的可行支架，ii) 生成基于肝脏 ECM 的成熟方案以生成肝细胞样细胞，以及 iii) 产生一种新的移植工程方法提供辅助肝脏支持。虽然这项工作利用肝脏作为模型器官，但这项工作的结果也将产生积极的影响，为未来复杂的器官工程技术奠定基础，该技术融合了几种不同的细胞类型，并可应用于其他器官（胰腺、肾脏等） .），并可能最终导致整个器官在体外发育。这里开发的 ESC 成熟方案预计将为干细胞工程领域做出重大贡献。脱细胞基质中的肝细胞培养也可能被证明是药物研究的新平台。公共卫生相关性：在美国，约有 3000 万人因不同原因患有肝脏疾病，美国每年约有 27,000 人因肝脏疾病死亡。目前，肝衰竭唯一确定的治疗方法是原位移植。然而，器官严重短缺，每年短缺约 3,000 个肝脏。类似的数字影响到大多数器官和组织，目前器官等待名单总数为 100,000 个请求，并且该数字每年以 5% 的速度增长。鉴于只有处于原始状态的器官才可以移植，原位移植的数量始终有限。这项研究的结果预计将通过开发一种利用干细胞衍生的肝细胞和废弃肝脏的天然基质进行辅助肝移植的新方法来直接改善公众健康，以治疗肝衰竭患者。