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
基本信息
- 批准号:10596894
- 负责人:
- 金额:$ 90.3万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 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:
- 关键词:AnimalsAsiaBiomedical EngineeringCardiac MyocytesCatalogsCattleCell Culture TechniquesCell CycleCellsCellular StructuresCollaborationsCollectionComplexCulture MediaCultured CellsCyclic GMPDataDependenceDevelopmentDevicesEncapsulatedEndotoxinsEnzyme-Linked Immunosorbent AssayEpithelial Cell ProliferationEpithelial Receptor CellEthicsEuropeExperimental DesignsFeedbackFetal ReductionFutureGoalsGrowthGrowth FactorHumanHydrogelsKnowledgeLifeManufacturerMarketingMeasuresMesenchymal Stem CellsMethodsMicrospheresModernizationPathway interactionsPhasePredictive FactorPreparationProductionProliferatingProteomicsRecombinant Growth FactorRecombinantsReproducibilityResearchRiskSamplingSerumSerum-Free Culture MediaSignal TransductionSmall Business Technology Transfer ResearchSourceStandardizationSterilityStimulusStructure of retinal pigment epitheliumSupporting CellT-LymphocyteTestingTimeViralWorkWritingcell growthcell replacement therapycell typeclinical lotcommercial launchcommercializationcontrolled releasefetal bovine serumhuman stem cellsimprovedmanufacturenerve stem cellpathogenpigment epithelium-derived factorresearch and developmentretinal stimulationstem cell replacementtherapy developmenttranscriptomics
项目摘要
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:
频繁使用胎牛血清(FBS)来支持细胞培养受到批间变异性、供应商
限制,病原体污染物和伦理问题。这些问题的重要性日益增加,
FBS在细胞研究和治疗开发中的新兴应用。FBS已被用于超过世纪,
培养细胞,但需要定义有效生产的培养条件,沿着现代
通过控制关键分子的递送的方法,现在可以解决与FBS使用相关的许多问题。
缓解减少动物产品的需求是至关重要的,使用人类生长因子的需求也是如此
来优化人类细胞的生长StemCultures公司专门从事控制生长因子(GF)
细胞培养基的成分。我们的目标是用生物工程DISCTM(定义插入物)取代FBS GF
用于可持续培养)装置,提供控释的人重组GF。高级细胞
制造DISCTM装置将解决有关FBS的问题并提高细胞生产的再现性。
在本申请中,我们将开发用于人视网膜色素上皮(RPE)细胞生长的DISCTM装置。
选择人RPE作为FBS替代的试验平台,因为这些细胞是干细胞的先锋
替代疗法在美国,欧洲和亚洲。减少FBS使用的一个障碍是缺乏以下知识:
FBS中对RPE细胞产物生长至关重要的GF含量。我们将测量FBS GF的水平,
刺激人RPE细胞增殖。选择的重组人GF将被封装在受控的-
释放GF微珠,随时间提供标准化的生长信号,以替代可变水平的
由FBS提供的具有确定水平的天然人重组GF的牛GF。GF微珠将
在析因设计实验中进行评估,以确定支持RPE细胞生长的微珠组合,
降低FBS水平。支持RPE细胞生长的微珠组合,无需FBS或在最低
水平的FBS将包装在可持续培养的定义插入物(DISCTM)装置中,以简化RPE细胞
制造。DISCTM将作为市售产品生产,以减少对FBS的依赖
商业上重要的RPE细胞。未来的工作将应用微珠/DISCTM平台策略,
降低对其他细胞类型的FBS依赖性。StemCultures(SCL)公司定期生产
微珠和DISCsTM用于在培养基和共定位神经干细胞中持续释放GF
研究所(NSCI)拥有广泛的RPE细胞培养专业知识。SCL-NSCI合作顺利完成
第一阶段的目标是使第二阶段的目标高度可行。
项目成果
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