Biotechnology Based Fetal Pulmonary Tissue Engineering

基于生物技术的胎儿肺组织工程

• 批准号: 7036852
• 负责人: CHRISTINE M FINCK
• 金额: $ 18.75万
• 依托单位: DREXEL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-09-07 至 2007-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7036852
• 关键词: biotechnology; cell morphology; cell population study; electron microscopy; embryo /fetus tissue /cell culture; gel; growth media; histology; immunocytochemistry; laboratory mouse; lung development; nanotechnology; perfusion; shear stress; tissue engineering; tissue support frame

DESCRIPTION (provided by applicant): Pulmonary hypoplasia or lung immaturity is found in as many as 15-20% of all neonatal autopsies. The ability to grow functional 3-dimensional lung tissue from cultured cells for eventual transplantation would be invaluable in preventing the neonatal mortality associated with pulmonary hypoplasia. The long term goal of this project is to alleviate neonatal pulmonary hypoplasia by augmenting damaged lung with in vitro engineered pulmonary tissue constructs. In order to achieve this goal, murine fetal pulmonary cells will be isolated, and utilizing engineered nano-based scaffolds, composite gels, growth factor enhanced media and an optimized microperfusion system, 3-dimensional lung constructs will be formed. The hypothesis of this project is that in an engineered micro-environment, cultured cells will be able to achieve cell to cell contact, adhere, proliferate, and differentiate into macroscopic 3-dimensional tissue constructs. The short term goal of this project is to systematically determine the parameters of a system for engineering lung tissue with appropriate morphology and cytodifferentiation. The specific aims of this project are to: 1- Optimize methods for isolating and culturing heterotypic populations of pulmonary fetal cells using enhanced culture media containing tissue-specific growth factors. 2- To facilitate adhesion, survival, growth and differentiation of cultured fetal lung cells over a prolonged period of time by growing these cells on organotypic 3-dimensional matrices composed of either composite gels or nanofibers electrospun from extracellular matrix derived molecules. 3- Utilize a microperfusion system to enable the continuous bathing of these 3-D constructs with nutrients as well as subject them to pulsatile flow induced shear forces. Morphology and cytodifferentiation of the ensuing 3-D contructs will be evaluated by histology, immunohistochemistry, electron microscopy and biochemical/ molecular biological techniques. The goal of this R-21 application is to explore and optimize the feasibility of this approach and to generate preliminary data for submission of a traditional R01 application.

描述（由申请人提供）：肺发育不全或肺不成熟在多达15-20%的新生儿尸检中发现。从培养的细胞中生长功能性三维肺组织用于最终移植的能力对于预防与肺发育不良相关的新生儿死亡率将是非常宝贵的。本项目的长期目标是通过体外工程化肺组织构建物增强受损肺来减轻新生儿肺发育不良。为了实现这一目标，将分离鼠胎肺细胞，并利用工程化的纳米基支架、复合凝胶、生长因子增强培养基和优化的微灌注系统，将形成三维肺构建体。该项目的假设是，在工程化的微环境中，培养的细胞将能够实现细胞与细胞的接触、粘附、增殖和分化成宏观的三维组织结构。本项目的短期目标是系统地确定具有适当形态和细胞分化的工程肺组织系统的参数。本项目的具体目标是：1-优化使用含有组织特异性生长因子的增强培养基分离和培养肺胎儿细胞异型群体的方法。2-通过使培养的胎肺细胞在由复合凝胶或由细胞外基质衍生分子电纺的纳米纤维组成的器官型三维基质上生长，促进这些细胞在延长的时间段内的粘附、存活、生长和分化。3-利用微灌注系统，使这些3-D结构与营养物连续沐浴，并使它们受到脉动流诱导的剪切力。通过组织学、免疫组织化学、电子显微镜和生物化学/分子生物学技术评价随后的三维结构的形态学和细胞分化。这项R-21申请的目的是探索和优化这种方法的可行性，并为提交传统的R 01申请生成初步数据。