Metabolomic Screening of Biomaterials for MSC Culture

用于 MSC 培养的生物材料的代谢组学筛选

• 批准号: 10396978
• 负责人: Johnna S Temenoff
• 金额: $ 20.67万
• 依托单位: GEORGIA INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-05-01 至 2024-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10396978
• 关键词: Biocompatible Materials; Biological; Biological Assay; Cell Aging; Cell Count; Cell Density; Cell Surface Receptors; Cell Therapy; Cells; Cellular Metabolic Process; Chronic; Chronic Disease; Clinical Trials; DNA; Data; Development; Disease; Early identification; Gene Expression; Goals; Growth Factor; Healthcare; Heparin; Human; Hydrogels; Integrins; Laboratories; Lead; Ligands; Liquid Chromatography; Maintenance; Mass Fragmentography; Mesenchymal; Metabolic; Metabolic Pathway; Metabolism; Methodology; Mission; Mitogens; Modeling; Modulus; Musculoskeletal; N-Cadherin; Outcome Measure; PF4 Gene; Pathology; Pathway interactions; Pattern; Phenotype; Process; Property; Protocols documentation; Public Health; Regenerative Medicine; Research; Sampling; Serum; Source; Stains; Stromal Cells; Styrenes; Sulfate; Surface; Testing; Therapeutic; Tissues; United States National Institutes of Health; Work; base; beta-Galactosidase; cost; density; design; experience; fitness; immunoregulation; improved; in vivo; innovation; metabolomics; paracrine; repaired; response; screening; senescence; stem; success; tissue culture; tissue repair; tool

PROJECT SUMMARY Mesenchymal stem/stromal cells (MSCs) have transformative healthcare potential, but to achieve therapeutic cell numbers, MSCs must be culture expanded. On tissue culture poly(styrene), however, MSC expansion is constrained by replicative senescence, and cells experience progressive loss of therapeutic potency with continued expansion. The lack of large numbers of reliably potent therapeutic cells is considered one of the most critical roadblocks to the success of MSC clinical trials. However, screening of improved MSC culture conditions has been hampered by the fact that there are no characterization tools that might allow prediction of cell fitness early in culture. Metabolism is the most dynamic level of cellular response, and metabolomics may enable the identification of early signatures associated with optimal cell product quality. Therefore, it is hypothesized that early metabolomics data from MSCs cultured on biomaterials will allow prediction of cell fitness later in culture and thus promote development of materials to promote a desired cell phenotype during expansion. The objective of this application is to determine the relationship between biomaterial carrier properties, cell metabolism, and maintenance of MSC fitness during expansion. This objective will be approached through the following specific aims: 1) Evaluate the effects of altering ligand type and stiffness of the biomaterial substrate on metabolism and replicative senescence of human MSCs during expansion in serum, and 2) Evaluate the effects of the substrate heparin content and pattern of initial seeding on metabolism and replicative senescence of human MSCs during expansion in serum-free conditions. The proposed work is innovative because it uses early cellular metabolic responses to design material substrates that will promote cell expansion without senescence in a range of media compositions. Results from these studies are expected to have an important positive impact because they will lead to more efficacious expansion protocols for MSCs and thus produce more effective cell-based therapies for a wide variety of diseases.

项目摘要 间充质干/基质细胞（MSC）具有变革性的医疗保健潜力，但要实现 治疗细胞数，MSC必须培养扩增。然而，在组织培养聚苯乙烯上， MSC扩增受到复制性衰老的限制，并且细胞经历细胞增殖的进行性丧失。 治疗效果持续扩大。缺乏大量可靠有效的治疗药物， 细胞被认为是MSC临床试验成功的最关键的障碍之一。然而，在这方面， 筛选改进的MSC培养条件受到以下事实的阻碍： 这是一种可能允许在培养早期预测细胞适应性的表征工具。新陈代谢是最重要的 动态水平的细胞反应和代谢组学可能使识别早期签名 与最佳细胞产品质量相关。因此，假设早期代谢组学数据 来自在生物材料上培养的MSC的生物相容性将允许预测培养中稍后的细胞适应性，从而促进 开发材料以在扩增期间促进所需的细胞表型。 本申请的目的是确定生物材料载体 性质、细胞代谢和在扩增期间维持MSC适应性。这一目标将是 通过以下具体目标来处理：1）评估改变配体类型的影响， 生物材料基质刚度对人骨髓间充质干细胞代谢和复制性衰老的影响 血清中扩增，和2）评价底物肝素含量和初始扩增模式的影响 接种对人骨髓间充质干细胞无血清扩增过程中代谢和复制衰老的影响 条件 这项拟议中的工作是创新的，因为它使用早期细胞代谢反应来设计材料 在一系列培养基组合物中促进细胞扩增而不衰老的基质。结果 这些研究预计将产生重要的积极影响，因为它们将导致更多 有效的扩增方案，从而产生更有效的细胞为基础的治疗广泛 各种疾病。