Molecular Engineering of Bioactive Hydrogels

生物活性水凝胶的分子工程

• 批准号: 7595085
• 负责人: DAVID V SCHAFFER
• 金额: $ 17.25万
• 依托单位: UNIVERSITY OF CALIFORNIA BERKELEY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-04-01 至 2011-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7595085
• 关键词: Address; Adverse effects; Affinity; Animals; Binding; Biological; Biology; Biomedical Engineering; Brain; Cell Count; Cell Culture System; Cell Differentiation process; Cell physiology; Cells; Clinical; Coculture Techniques; Collection; Complex; Conditioned Culture Media; Congestive Heart Failure; Development; Diabetes Mellitus; Disease; Engineering; Engraftment; Environment; Epitopes; Extracellular Matrix; Extracellular Matrix Proteins; Goals; Grant; Heart; Human; Hydrogels; Immune; In Vitro; Injury; Ligands; Liver; Medicine; Methods; Molecular; Molecular Biology; Mus; Muscle; Pancreas; Parkinson Disease; Patients; Peptides; Process; Proteins; Regenerative Medicine; Reproducibility; Science; Serum; Signal Transduction; Source; Stem cells; Surface; System; Technology; Tissue Engineering; Tissues; Treatment Efficacy; base; bone; cell behavior; cell type; human embryonic stem cell; immunogenic; in vivo; large scale production; leukemia; novel; pathogen; public health relevance; reconstruction; scale up; self-renewal; stem cell biology; stem cell population; synthetic peptide; therapy design; transmission process

Description (provided by applicant): Human embryonic stem cells (hESCs) have strong potential as sources of cells for the treatment for disease and injury (e.g. tissue engineering and reconstruction, diabetes, Parkinson's Disease, leukemia, congestive heart failure, etc.). The successful integration of hESC into such therapies will hinge upon three critical steps: their expansion without differentiation (i.e., self-renewal), their differentiation into a specific cell type or collection of cell types, and the promotion of their survival and functional integration into existing tissue. However, controlling cell behavior during each of these steps will require precise control over the cellular microenvironment. This poses a major challenge ex vivo in current hESC culture systems, which range from co-culture with feeder cells to serum-free systems where cells are cultured on complex extracellular matrix proteins. All such systems involve animal or human proteins, which pose problems for pathogen transmission, immune rejection, limited reproducibility, and scale up to a clinical process. To achieve the intended goals of regenerative medicine, methods for the precise control of the survival, proliferation, and differentiation of stem cell populations in vitro and in vivo are necessary. Here, we propose to develop a completely synthetic environment to precisely control hESC self-renewal in culture. Specifically, we will engineer a tunable and well-defined environment presenting a completely "synthetic extracellular matrix" (ECM) and chemically-defined media to control the self-renewal/expansion of hESCs. Furthermore, we will develop high throughput approaches to identify synthetic peptide ligands for functionalization to the synthetic ECM and promotion of hESC self-renewal. If hESCs can be derived and maintained within this fully synthetic microenvironment, then it will be possible to eliminate pathogen transmission associated with mouse or human feeder layers, provide a scalable basis for large-scale production of hESCs, and provide a precise base for further development to control hES cell differentiation. Furthermore, the result will be a technology platform that can be generally applied to numerous stem cell populations and used to investigate the basic biological/developmental mechanisms underlying self-renewal. Public Health Relevance: The development of novel, bioactive materials has significant potential for exerting precise control over cell function, both for fundamental biological studies and applications in tissue engineering and regenerative medicine. For example, developing synthetic, bioactive material systems to promote the self-renewal and expansion of human embryonic stem cells will have numerous biomedical applications including the design of therapies for disease or injury in the muscle, bone, brain, heart, liver, pancreas, and other tissues. The novel blend of stem cell biology, materials science, molecular biology, and bioengineering described in this proposal will be well suited to addressing an important problem, i.e. stem cell control, at the interface of biology, engineering, and medicine

描述（由申请人提供）：人类胚胎干细胞（hESC）作为治疗疾病和损伤（例如组织工程和重建、糖尿病、帕金森病、白血病、充血性心力衰竭等）的细胞来源具有强大的潜力。 hESC 成功整合到此类疗法中将取决于三个关键步骤：它们在不分化的情况下扩增（即自我更新），它们分化为特定细胞类型或细胞类型集合，以及促进它们的存活和功能整合现有组织。然而，在每个步骤中控制细胞行为都需要精确控制细胞微环境。这对当前 hESC 培养系统的离体提出了重大挑战，这些系统的范围从与饲养细胞共培养到细胞在复杂的细胞外基质蛋白上培养的无血清系统。所有此类系统都涉及动物或人类蛋白质，这会给病原体传播、免疫排斥、有限的再现性以及扩大到临床过程带来问题。为了实现再生医学的预期目标，需要在体外​​和体内精确控制干细胞群的存活、增殖和分化的方法。在这里，我们建议开发一个完全合成的环境来精确控制培养中 hESC 的自我更新。具体来说，我们将设计一个可调节且明确的环境，提供完全“合成的细胞外基质”（ECM）和化学成分确定的培养基，以控制 hESC 的自我更新/扩张。此外，我们将 开发高通量方法来识别合成肽配体，以对合成 ECM 进行功能化并促进 hESC 自我更新。如果hESCs能够在这种完全合成的微环境中衍生和维持，那么将有可能消除与小鼠或人类饲养层相关的病原体传播，为hESCs的大规模生产提供可扩展的基础，并为进一步开发提供精确的基础控制 hES 细胞分化。此外，其结果将是一个可普遍应用于众多干细胞群体并用于研究自我更新的基本生物/发育机制的技术平台。公共健康相关性：新型生物活性材料的开发在对细胞功能进行精确控制方面具有巨大潜力，无论是对于基础生物学研究还是组织工程和再生医学的应用。例如，开发合成的生物活性材料系统来促进人类胚胎干细胞的自我更新和扩增将具有许多生物医学应用，包括设计针对肌肉、骨骼、大脑、心脏、肝脏、胰腺等疾病或损伤的疗法，和其他组织。该提案中描述的干细胞生物学、材料科学、分子生物学和生物工程的新颖融合将非常适合解决生物学、工程和医学交叉领域的一个重要问题，即干细胞控制

项目成果

Surface creasing instability of soft polyacrylamide cell culture substrates.

软聚丙烯酰胺细胞培养基质的表面折痕不稳定性。

• DOI: 10.1016/j.bpj.2010.09.045
• 发表时间: 2010-12-15
• 期刊: Biophysical journal
• 影响因子: 3.4
• 作者: Krishanu Saha;Jungwook Kim;E. Irwin;Jinhwan Yoon;F. Momin;Verónica Trujillo;D. Schaffer;K. Healy;R. Hayward
• 通讯作者: R. Hayward

Exploiting bacterial peptide display technology to engineer biomaterials for neural stem cell culture.

利用细菌肽展示技术来设计用于神经干细胞培养的生物材料。

• 发表时间: 2011-02
• 影响因子: 14
• 作者: Little, Lauren E;Dane, Karen Y;Daugherty, Patrick S;Healy, Kevin E;Schaffer, David V
• 通讯作者: Schaffer, David V

Engineering strategies to emulate the stem cell niche.

模拟干细胞生态位的工程策略。

• DOI: 10.1016/j.tibtech.2009.11.008
• 发表时间: 2010-03-01
• 期刊: Trends in biotechnology
• 影响因子: 17.3
• 作者: T. Vazin;D. Schaffer
• 通讯作者: D. Schaffer

Characterization of integrin engagement during defined human embryonic stem cell culture.

确定的人胚胎干细胞培养过程中整合素参与的表征。

• 发表时间: 2010-04
• 作者: Meng, Ying;Eshghi, Shawdee;Li, Ying J;Schmidt, Ray;Schaffer, David V;Healy, Kevin E
• 通讯作者: Healy, Kevin E

Neural stem cell adhesion and proliferation on phospholipid bilayers functionalized with RGD peptides.

使用 RGD 肽功能化的磷脂双层上的神经干细胞粘附和增殖。

• DOI: 10.1016/j.biomaterials.2010.07.104
• 发表时间: 2010-11
• 期刊: Biomaterials
• 影响因子: 14
• 作者: Ananthanarayanan, Badriprasad;Little, Lauren;Schaffer, David V.;Healy, Kevin E.;Tirrell, Matthew
• 通讯作者: Tirrell, Matthew