Craniofacial tissue engineering with human embryonic an*

人类胚胎颅面组织工程*

• 批准号: 7035847
• 负责人: ROBERT Samuel LANGER
• 金额: $ 37.91万
• 依托单位: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-04-01 至 2010-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7035847
• 关键词: bioengineering /biomedical engineering; biomaterial interface interaction; biotechnology; cell differentiation; cell morphology; cell proliferation; chondrocytes; craniofacial; gel; gene expression; high throughput technology; human embryonic stem cell line; immunocytochemistry; laboratory rat; microarray technology; musculoskeletal regeneration; osteoblasts; polymerase chain reaction; polymers; tissue engineering; tissue support frame

DESCRIPTION (provided by applicant): We propose to use our established and well documented history in the development of biomaterials as scaffolding for tissue replacement as a platform for stem cell approaches in craniofacial tissue regeneration. The governing hypotheses of this work include: i) that human stem cells of embryonic origin can be differentiated into craniofacial tissue producing cells such as osteoblasts and chondrocytes, ii) that highthroughput techniques can be used for the screening of large numbers of media and polymer candidates for potential use with tissue engineering scaffolds, and iii) that the appropriate combination of stem cells with 3-dimensional polymer scaffolds can be used for the production of complex craniofacial tissues. To test these hypotheses, we propose the following: Aim 1: Examine the in vitro differentiation of human embryonic stem cells into precursors of craniofacial tissues. Our previous work investigating the in vitro differentiation of human embryonic stem cells into various cell types and tissues will be continued to assess conditions necessary for differentiation into cell types necessary for craniofacial tissue formation. Aim 2: Investigate the interaction of human stem cells with polymer surfaces using high-throughput screening technology. Recently, we have developed techniques for the rapid screening of cell/polymer interactions using nanoliter-scale microarrays of various monomers and polymers. Success will be assessed through cell proliferation and the presence of tissue specific markers for craniofacial tissues. Aim 3: Assess in vitro tissue production by differentiated and undifferentiated human embryonic stem cell seeded polymeric scaffolds incorporating various genes and growth factors. Polymeric materials that support the differentiation of human stem cells into craniofacial tissues will be fabricated into 3-dimensional scaffolds incorporating various genes and growth factors and analyzed by histological staining of various markers. Aim 4: Optimal candidates from the previous aims will be seeded with differentiated and undifferentiated cells and implanted either subcutaneously or in a critical sized defect model in rats. The subcutaneous model in athymic rats will be used to assess tissue production by stem cell seeded scaffolds in an in vivo environment, including scaffolds with cells differentiated into multiple phenotypes. The cranial defect model in athymic rats is a clinically relevant model that is commonly used as a measure of bone regeneration.

描述（由申请人提供）：我们建议利用我们在生物材料开发中既定且有据可查的历史作为组织替代的支架，作为颅面组织再生干细胞方法的平台。这项工作的主导假设包括：i）胚胎来源的人类干细胞可以分化为颅面组织生成细胞，如成骨细胞和软骨细胞，ii）高通量技术可用于筛选大量培养基和聚合物候选物与组织工程支架的潜在用途，以及iii）干细胞与3维聚合物支架的适当组合可用于生产复杂的颅面组织。为了检验这些假设，我们提出以下建议： 目标 1：检查人胚胎干细胞体外分化为颅面组织前体的情况。我们之前研究人类胚胎干细胞体外分化为各种细胞类型和组织的工作将继续评估分化为颅面组织形成所需的细胞类型所需的条件。 目标 2：利用高通量筛选技术研究人类干细胞与聚合物表面的相互作用。最近，我们开发了使用各种单体和聚合物的纳升级微阵列快速筛选细胞/聚合物相互作用的技术。将通过细胞增殖和颅面组织组织特异性标记物的存在来评估成功。目标 3：评估含有各种基因和生长因子的分化和未分化人胚胎干细胞接种的聚合物支架的体外组织生产。支持人类干细胞分化为颅面组织的聚合物材料将被制造成包含各种基因和生长因子的3维支架，并通过各种标记的组织学染色进行分析。目标 4：先前目标的最佳候选者将被接种分化和未分化细胞，并植入皮下或大鼠的临界尺寸缺损模型中。无胸腺大鼠的皮下模型将用于评估体内环境中干细胞接种支架的组织生产，包括细胞分化为多种表型的支架。无胸腺大鼠的颅骨缺损模型是一种临床相关模型，通常用作骨再生的测量。