Biotechnology Based Fetal Pulmonary Tissue Engineering

基于生物技术的胎儿肺组织工程

• 批准号: 7140671
• 负责人: CHRISTINE M FINCK
• 金额: $ 21.97万
• 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/7140671
• 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）：在所有新生儿尸检中，多达 15-20% 的人发现肺发育不全或肺不成熟。从培养细胞中培养出功能性三维肺组织以供最终移植的能力对于预防与肺发育不全相关的新生儿死亡具有不可估量的价值。该项目的长期目标是通过体外工程肺组织结构增强受损的肺来缓解新生儿肺发育不全。为了实现这一目标，将分离小鼠胎儿肺细胞，并利用工程纳米支架、复合凝胶、生长因子增强培养基和优化的微灌注系统，形成 3 维肺结构。该项目的假设是，在工程微环境中，培养的细胞将能够实现细胞与细胞的接触、粘附、增殖并分化成宏观的 3 维组织结构。该项目的短期目标是系统地确定用于工程化具有适当形态和细胞分化的肺组织的系统的参数。该项目的具体目标是： 1- 使用含有组织特异性生长因子的增强培养基来优化分离和培养肺胎儿细胞异型群的方法。 2- 通过在由复合凝胶或由细胞外基质衍生分子静电纺丝而成的纳米纤维组成的器官型 3 维基质上培养这些细胞，促进培养的胎儿肺细胞在较长时间内的粘附、存活、生长和分化。 3- 利用微灌注系统使这些 3-D 结构能够连续沐浴营养物质，并使其承受脉动流引起的剪切力。随后的 3-D 结构的形态学和细胞分化将通过组织学、免疫组织化学、电子显微镜和生物化学/分子生物学技术进行评估。此 R-21 申请的目标是探索和优化该方法的可行性，并生成用于提交传统 R01 申请的初步数据。