Controlled-release Microbeads to Replace Growth Factors in Fetal Bovine Serum

控释微珠替代胎牛血清中的生长因子

• 批准号: 10596894
• 负责人: Kristina Roberts
• 金额: $ 90.3万
• 依托单位: STEMCULTURES, LLC
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-01 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10596894
• 关键词: Animals; Asia; Biomedical Engineering; Cardiac Myocytes; Catalogs; Cattle; Cell Culture Techniques; Cell Cycle; Cells; Cellular Structures; Collaborations; Collection; Complex; Culture Media; Cultured Cells; Cyclic GMP; Data; Dependence; Development; Devices; Encapsulated; Endotoxins; Enzyme-Linked Immunosorbent Assay; Epithelial Cell Proliferation; Epithelial Receptor Cell; Ethics; Europe; Experimental Designs; Feedback; Fetal Reduction; Future; Goals; Growth; Growth Factor; Human; Hydrogels; Knowledge; Life; Manufacturer; Marketing; Measures; Mesenchymal Stem Cells; Methods; Microspheres; Modernization; Pathway interactions; Phase; Predictive Factor; Preparation; Production; Proliferating; Proteomics; Recombinant Growth Factor; Recombinants; Reproducibility; Research; Risk; Sampling; Serum; Serum-Free Culture Media; Signal Transduction; Small Business Technology Transfer Research; Source; Standardization; Sterility; Stimulus; Structure of retinal pigment epithelium; Supporting Cell; T-Lymphocyte; Testing; Time; Viral; Work; Writing; cell growth; cell replacement therapy; cell type; clinical lot; commercial launch; commercialization; controlled release; fetal bovine serum; human stem cells; improved; manufacture; nerve stem cell; pathogen; pigment epithelium-derived factor; research and development; retinal stimulation; stem cell replacement; therapy development; transcriptomics

Scientific Abstract: The frequent use of fetal bovine serum (FBS) to support cell cultures is challenged by lot-to-lot variability, supply constraints, pathogen contaminants and ethical questions. These issues have increased significance due to the burgeoning use of FBS in cell research and therapy development. FBS has been used for over a century to culture cells, but with the need to define culture conditions for efficient manufacturing, along with modern approaches to control delivery of key molecules, many of the problems associated with FBS use can now be alleviated. The need to reduce animal products is critically important, as is the need to use human growth factors to optimize human cell growth. The company StemCultures specializes in controlling the growth factor (GF) component of cell culture media. Our goal is to replace FBS GFs with a bioengineered DISCTM (Defined Insert for Sustainable Culture) device providing controlled-release human recombinant GFs. The advanced cell manufacturing DISCTM device will address concerns about FBS and improve the reproducibility of cell production. In this application we will develop a DISCTM device for growth of human retinal pigment epithelial (RPE) cells. Human RPE was selected as a testbed for FBS replacement as these cells are a vanguard for stem cell replacement therapy in the US, Europe and Asia. A barrier to reducing FBS use is the lack of knowledge about the GF content of FBS that is critical for growth of the RPE cell product. We will measure levels of FBS GFs that stimulate human RPE cell proliferation. Selected recombinant human GFs will be encapsulated in controlled- release GF microbeads that provide a standardized growth signal over time, to replace the variable levels of bovine GFs provided by FBS with defined levels of native human recombinant GFs. The GF microbeads will be assessed in a factorial design experiment to identify microbead combinations that support RPE cell growth at a reduced level of FBS. The microbead combination that supports RPE cell growth without FBS or at the lowest level of FBS will be packaged in a Defined Insert for Sustainable Culture (DISCTM) device to simplify RPE cell manufacture. The DISCTM will be manufactured as a commercial product marketed to reduce FBS dependence of the commercially important RPE cells. Future work will apply the microbead/DISCTM platform strategy to reduce FBS dependence of other cell types. The company StemCultures (SCL) routinely manufactures microbeads and DISCsTM for sustained release of GFs in culture media and the co-located Neural Stem Cell Institute (NSCI) has extensive RPE cell culture expertise. The SCL-NSCI collaboration successfully completed Phase 1 aims, in preparation to make Phase 2 goals highly feasible.

科学摘要： 经常使用胎牛血清（FBS）来支持细胞培养物，这是由洛特变异性挑战 约束，病原体污染物和道德问题。由于 迅速使用FBS在细胞研究和治疗开发中。 FB已有一个多世纪 培养细胞，但需要定义有效制造的培养条件，以及现代 控制关键分子传递的方法，现在可以使用FBS的许多问题 缓解。减少动物产品的需求至关重要，需要使用人类生长因素 优化人类细胞生长。该公司的干燥文化专门控制增长因子（GF） 细胞培养基的组成部分。我们的目标是用生物工程的拆卸替换FBS GFS（定义的插入物 对于可持续文化）设备提供受控释放的人类重组GF。高级单元格 制造DISCTM设备将解决对FBS的担忧，并提高细胞生产的可重复性。 在此应用中，我们将开发一种用于人类视网膜色素上皮（RPE）细胞生长的DISCTM设备。 选择人RPE作为FBS替代的测试床，因为这些细胞是干细胞的先锋 美国，欧洲和亚洲的替代疗法。减少FBS使用的障碍是缺乏有关 FBS的GF含量对于RPE细胞产物的生长至关重要。我们将测量FBS GF的水平 刺激人RPE细胞增殖。选定的重组人GF将被封装在受控 - 释放GF Microbeads，随着时间的推移提供标准化的增长信号，以取代可变水平 FBS提供的牛GFs具有定义的本地人类重组GF。 GF Microbeads将是 在阶乘设计实验中进行了评估，以鉴定支持RPE细胞生长的微粒组合 FBS的水平降低。支持无FB或最低的RPE细胞生长的Microbead组合 FB的水平将包装在可持续文化（DISCTM）设备的定义插入物中，以简化RPE单元格 生产。该拆除将作为销售的商业产品制造，以减少FBS依赖性 商业上重要的RPE细胞。未来的工作将应用Microbead/Disctm平台策略 减少其他细胞类型的FBS依赖性。公司干燥文化（SCL）通常制造 Microbeads和Discstm，用于在培养基中持续释放GFS和共同置换的神经干细胞 Institute（NSCI）具有广泛的RPE细胞培养专业知识。 SCL-NSCI合作成功完成 第1阶段的目的是准备实现高度可行的2阶段目标。