CORE--CELL CULTURE

核心--细胞培养

• 批准号: 6323265
• 负责人: JAMES RHEINWALD
• 金额: $ 7.67万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-04-01 至 2001-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6323265
• 关键词: animal tissue; biomedical facility; fibroblasts; growth media; human tissue; keratinocyte; melanocyte; skin; tissue /cell culture; transfection; vascular endothelium

In contrast to most mammalian organs, many cellular elements of normal human skin can now be cultured. Techniques to culture human keratinocytes (first evolved by HSDRC member Dr. Green), dermal fibroblasts, melanocytes, and microvascular endothelial cells are now standardized, and most of these cells can be obtained from at least one commercial vendor (e.g., Clonetics). Each cell type requires different medium, however, and it is impractical for one laboratory to grow all such cells. Furthermore, unless study of one of these individual cell in a consistent and detailed fashion, it is impractical to initiate the culture of such cells on a regular basis. The main goal of the Cell Culture core is to facilitate experimental use of freshly cultured skin cells, of both human and murine origin. With regards to murine cells, particular emphasis will be placed on the culturing of transgenic-derived keratinocytes, melanocytes, dermal fibroblasts and endothelial cells. The specific aims of the proposal are to provide the following services: 1) Procurement of human and murine skin. 2) Special media preparation for specific cell types. 3) Isolation and cultivation of skin-derived primary cells, including large scale preparations for biochemical studies (such as RNA and cDNA library preparation; cytokine and enzyme purifications, etc.). 4) Training in the in vitro isolation of cellular elements of skin, such that investigators appropriately trained and having access to appropriate media can initiate and maintain cell cultures in their laboratories.

与大多数哺乳动物器官相比，正常的许多细胞元件 现在可以培养人类皮肤。人角质形成细胞培养技术 （首先由HSDRC成员Green博士进化而来），真皮成纤维细胞， 黑素细胞和微血管内皮细胞现已标准化，并且 大多数这些电池可以从至少一个商业供应商处获得 （例如克隆学）。然而，每种细胞类型需要不同的培养基 一个实验室培养所有此类细胞是不切实际的。此外， 除非对这些单个细胞之一进行一致和详细的研究 时尚，在 定期。 细胞培养核心的主要目标是促进实验使用 新鲜培养的人类和小鼠皮肤细胞。和 关于小鼠细胞，特别强调 转基因衍生的角质形成细胞、黑素细胞、真皮细胞的培养 成纤维细胞和内皮细胞。 该提案的具体目标是提供以下服务： 1) 采购人和鼠皮肤。 2) 针对特定细胞类型的特殊培养基制备。 3）皮肤来源原代细胞的分离和培养，包括 用于生化研究的大规模制剂（例如 RNA 和 cDNA 文库准备；细胞因子和酶纯化等）。 4) 皮肤细胞成分的体外分离培训，例如 调查人员经过适当培训并能够获得适当的信息 培养基可以在实验室中启动和维持细胞培养。