CORE--CELL CULTURE/TISSUE PREPARATION

核心——细胞培养/组织制备

• 批准号: 6197177
• 负责人: MARY L WILLIAMS
• 金额: $ 19.83万
• 依托单位: NORTHERN CALIFORNIA INSTITUTE/RES/EDU
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-08-01 至 2000-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6197177
• 关键词: biomedical facility; keratinocyte; tissue /cell culture

The Cell Culture Core supplies high quality cells for the Program Project investigators. The facility is maintained such that it meets and exceeds the relevant federal health and safety standards. The majority of cells provided by the facility are foreskin-derived normal human keratinocytes. These cells are plated with media, conditions, density etc. as required by each program project investigator. Presently the facility is providing over 250 million cells per week to investigators, but it can produce greater than or equal to 500 million per week on demand. In addition, the facility has a large library of transformed and genetically altered lines that are available to investigators. The most frequently used by the investigators are the squamous cell carcinoma lines (SCC-4, SCC-12F2, SCC-12B2); and low passage cells (HaCaT, A431). These are maintained at low levels which allows for rapid boosting of cell numbers at the investigator's request. Dr. Mary L. Williams, Director, & Ms. Sally Pennypacker, Heady Technician, not only provide in vitro culture knowledge and expertise for other investigators, but also insure that health and safety inspections are passed; hoods and incubators are tested and serviced regularly; and that cultures are tested monthly for mycoplasma. Both Dr. Williams & Ms. Pennypacker also possess expertise in culturing basal cell carcinoma cells, adult skin keratinocytes and fibroblasts, fetal skin keratinocytes, late passage keratinocytes, fetal mouse skin keratinocytes; newborn and adult mouse keratinocytes; lifted keratinocyte cultures; and proficiency in explant methods. Core A also is developing fibroblast-free fetal keratinocytes, and optimizing both BCC isolation and late-passage keratinocyte cultures (i.e., greater pass 25). A fundamental part of the operation of Core A is the interaction between its personnel and members of the Program Project, allowing researchers to obtain high-quality cultures specifically tailored to their needs, while providing direct feedback for optimizing of techniques and protocols. The facility is now equipped with amber lights for work with light-sensitive compounds (e.g., retinoids, vitamin D and derivatives). Finally, Core A provides both maximum numbers of high quality cells and advice on in vitro methods to investigators, who rely heavily on this facility. The continued production of healthy keratinocytes will remain the priority of this key facility.

细胞培养核心为项目研究人员提供高质量的细胞。该设施的维护符合并超过相关的联邦健康和安全标准。该机构提供的大多数细胞是包皮衍生的正常人角质形成细胞。根据每个项目研究者的要求，将这些细胞用培养基、条件、密度等进行平板培养。目前，该设施每周向调查人员提供超过2.5亿个细胞，但每周可按需生产超过或等于5亿个细胞。此外，该设施还有一个大型的转化和遗传改变系库，可供研究人员使用。研究者最常使用的是鳞状细胞癌细胞系（SCC-4、SCC-12 F2、SCC-12 B2）和低传代细胞（HaCaT、A431）。这些维持在低水平，允许在研究者的要求下快速增加细胞数量。玛丽博士主任威廉姆斯和高级技术员萨莉·彭尼帕克女士不仅为其他研究人员提供体外培养知识和专业知识，而且还确保通过健康和安全检查;定期测试和维修通风橱和培养箱;每月对培养物进行支原体测试。威廉姆斯博士和Pennypacker女士还拥有培养基底细胞癌细胞、成人皮肤角质形成细胞和成纤维细胞、胎儿皮肤角质形成细胞、晚期传代角质形成细胞、胎鼠皮肤角质形成细胞、新生和成年小鼠角质形成细胞、提升角质形成细胞培养物的专业知识;并精通外植体方法。核心A也正在开发无成纤维细胞的胎儿角质形成细胞，并优化BCC分离和晚期传代角质形成细胞培养物（即，更大的通行证25）。核心A运作的一个基本部分是其工作人员与项目成员之间的互动，使研究人员能够获得专门针对其需求的高质量培养物，同时为优化技术和方案提供直接反馈。该设施现在配备了琥珀色灯，用于处理光敏化合物（例如，类维生素A、维生素D和衍生物）。最后，核心A为严重依赖该设施的研究人员提供最大数量的高质量细胞和体外方法的建议。健康角质细胞的持续生产将仍然是这个关键设施的优先事项。