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Organoid culture media formulated with growth factors of defined cellular activity

机译：用定义的细胞活性的生长因子配制的类器官培养基

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The media formulations necessary for deriving and sustaining organoids from epithelial tissues such as prostate, colon, gastric, liver, pancreas, and others have been established. Critical components of organoid media are a set of growth factors that include R-spondins and BMP signalling antagonists such as Noggin or Gremlin 1. Currently, the practical limitations for formulating organoid media of reproducible potency and larger-scale media production that have hampered further technological applications of organoid technology include: the cost of growth factors such as R-spondins and Gremlin 1/Noggin and their production as defined specific activities free of contaminants that may affect organoid growth. Here we report the production of highly pure recombinant Gremlin 1 and R-spondin 1 from bacterial expression for use in organoid media. We detail the workflow for Gremlin 1 and R-spondin 1 expression, purification, quantification of cellular activity, quality control and use in media formulated for culturing organoids derived from a number of tissues. The development of precisely formulated, cost-effective media of defined specific activity will engender the development of novel applications for organoid technology.
机译：已经建立了从上皮组织例如前列腺，结肠，胃，肝，胰腺和其他组织衍生和维持类器官所需的培养基配方。类器官介质的关键成分是一组生长因子，包括R-spondins和BMP信号拮抗剂，例如Noggin或Gremlin1。目前，配制具有可重复效价的类器官介质的实际限制和大规模培养基生产已阻碍了进一步的技术发展。类器官技术的应用包括：生长因子（例如R-spondins和Gremlin 1 / Noggin）的成本及其定义的特定活动的生产，这些活动不含可能影响类器官生长的污染物。在这里，我们报告了从细菌表达用于类器官培养基的高纯度重组Gremlin 1和R-spondin 1的生产。我们详细介绍了Gremlin 1和R-spondin 1的表达，纯化，细胞活性定量，质量控制以及在配制用于培养源自许多组织的类器官的培养基中的使用的工作流程。精确配制的，具有特定比活的高性价比介质的开发将促使类器官技术的新应用程序的开发。

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