CORE--CELL CULTURE

核心--细胞培养

• 批准号: 6478144
• 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: 2001
• 资助国家: 美国
• 起止时间: 2001-04-01 至 2002-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6478144
• 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反，许多正常的细胞元素 现在可以培养人的皮肤。培养人角质形成细胞的技术 （首先由HSDRC成员Green Dr. Green演变），皮肤成纤维细胞， 黑素细胞和微血管内皮细胞现在已标准化，并且 这些单元中的大多数可以从至少一个商业供应商那里获得 （例如，克隆学）。但是，每种细胞类型都需要不同的介质，并且 一个实验室种植所有此类细胞是不切实际的。此外， 除非在一致且详细地研究这些单个单元之一 时尚，在A上启动此类细胞的培养是不切实际的 定期。 细胞培养核心的主要目标是促进实验使用 人类和鼠起源的新鲜培养的皮肤细胞。和 关于鼠类细胞的问候，将特别重点放在 培养转基因的角质形成细胞，黑素细胞，真皮 成纤维细胞和内皮细胞。 该提案的具体目的是提供以下服务： 1）采购人和鼠皮。 2）特定细胞类型的特殊媒体准备。 3）皮肤来源的原代细胞的隔离和培养，包括 生化研究的大规模制备（例如RNA和cDNA 图书馆准备；细胞因子和酶纯化等）。 4）训练皮肤细胞元素的体外分离， 调查人员接受了适当的培训并获得适当的访问权限 媒体可以在其实验室中启动和维护细胞培养物。