Engineering of hepatic grafts with decellularized native matrices

描述（由申请人提供）： 这是索托-古铁雷斯博士的职业资助申请，他是医学博士[sic]博士专门研究细胞疗法和肝病索托-古铁雷斯博士最近开发了将干细胞分化为肝细胞的新方法，他的兴趣已经转移到将这些工具应用于器官工程。为了使自己成为器官工程领域的独立研究人员，索托-古铁雷斯博士在哈佛医学院外科和生物工程学海伦·安德鲁斯·本尼迪克特教授和马萨诸塞州总医院医学工程中心主任马丁·亚穆什博士的赞助下提交了这份五年职业发展计划，其中包括i）器官工程学的强化实践培训，分离的肝基质灌注和干细胞分化，ii）学术课程和研讨会，以及iii）选定咨询委员会的指导。我们的长期目标是开发一个天然的肝脏平台，用于干细胞衍生的肝细胞的完全和稳定成熟，并设计功能性肝移植物。本研究的目的是开发一种体外灌注肝基质支架系统，用于干细胞肝源性细胞的再增殖和肝成熟，并研究其在肝功能受损患者中用于建立辅助肝移植物的就业。这里要测试的中心假设是，从丢弃的肝脏中获得的脱细胞化的天然肝脏支架可以被广泛地重新填充，并且可以为干细胞衍生的肝细胞提供足够的成熟环境，并且这种器官培养系统可以用于设计用于移植的代谢功能性肝移植物。该研究的基本原理是，化学损伤，不可移植的器官数量远远超过目前的需求，它们为基于干细胞的治疗提供了一个理想的平台。本文所述的工作预计i）建立脱细胞基质中肝细胞再细胞化和植入的方案，ii）产生干细胞衍生肝细胞的再增殖和成熟系统，以及iii）开发一种新的肝移植工程方法，用于肝脏疾病中作为辅助肝脏支持的移植。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建立基础和平台，这些技术结合了几种不同的细胞类型，可以应用于其他器官，并可能导致整个器官的体外发育。 公共卫生相关性：在美国，肝脏疾病每年导致约27，000人死亡，其中2，500人在等待肝脏移植的名单上死亡。肝脏相关死亡的一个主要原因是有限的供体库。该项目旨在通过利用目前废弃的供体肝脏作为干细胞衍生的肝细胞的天然支架来构成可成功移植的肝脏移植物，以改善公共健康。

DESCRIPTION (provided by applicant): This is a career grant application for Dr. Soto-Gutierrez; an M.D.; [sic] Ph.D. who specializes in cell therapies and liver diseases. Dr. Soto-Gutierrez has recently developed novel methods of differentiating stem cells into liver cells, and his interests have shifted to application of such tools to organ engineering. To establish himself as an independent researcher in the field of organ engineering, Dr. Soto-Gutierrez submits this five-year career development plan under the sponsorship of Dr. Martin Yarmush, the Helen Andrus Benedict Professor of Surgery and Bioengineering at Harvard Medical School and the director of the Center for Engineering in Medicine at the Massachusetts General Hospital, which includes i) intensive hand-on training in organ engineering, isolated liver-matrices perfusion and stem cell differentiation, ii) academic courses and seminars, and iii) guidance of a select advisory committee. Our long-term goals are to develop a natural hepatic platform for complete and stable maturation of stem-derived liver cells and to engineer functional liver grafts. The objectives of the proposed study are to develop an extracorporeal perfusion liver-matrix-scaffold system for repopulation and hepatic maturation of stem liver-derived cells, and investigate its employment for the creation of auxiliary liver grafts for transplantation in patients with impaired hepatic function. The central hypothesis to be tested here is that the decellularized natural liver scaffold, obtained from discarded livers, can be extensively repopulated and can provide an adequate maturation environment for stem derived liver cells, and this organ culture system can be employed to engineer metabolic functional liver grafts for transplantation. The rationale of the study is that is chemically damaged, untransplantable organs are available in numbers that far surpass the current demand, and they present an ideal platform for delivery for stem cell-based therapies. The work described here is expected to i) establish a protocol for recellularization and engraftment of hepatocytes [sic] in decellularized matrices, ii) generate a repopulation and maturation system for stem cell-derived hepatocytes, and iii) develop a novel method of liver graft engineering for transplantation as auxiliary liver support in hepatic diseases. The results of this work will also have a positive impact by establishing the basis and platform for future sophisticated organ engineering techniques that incorporates several different cell types, can be applied to other organs and may lead to development of entire organs in vitro. PUBLIC HEALTH RELEVANCE: Liver diseases cause about 27,000 deaths annually in the US, 2,500 of which perish on the waiting list for liver transplant. A major reason of liver related deaths is the limited donor pool. This project aims to improve public health by utilizing currently discarded donor livers as natural scaffolds for stem cell-derived liver cells to constitute livers grafts to be transplantable with success.