CORE-- CELL CULTURE

核心--细胞培养

• 批准号: 6597607
• 负责人: Thomas A Kocarek
• 金额: $ 17.41万
• 依托单位: WAYNE STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-04-01 至 2003-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6597607
• 关键词: antiseptic sterilization; biomedical facility; environmental toxicology; isozymes; karyotype; restriction fragment length polymorphism; tissue /cell culture; tissue /cell preparation

The Cell Culture Facility Core (CCFC) is housed in two laboratory rooms in the Institute of Chemical Toxicology, that contain all items of equipment necessary for cell culture work. Another laboratory for viral transfection of cultured cells has been established within the Wayne State University School of Medicine, but it is not clear to what degree this latter lab is under the control of the CCFC. Dr. Thomas Kocarek is the core Leader and is requesting 20% of his salary for this role. He is assisted by two full time technicians. The facility provides the following services: 1) preparation of primary cultures of animal hepatocytes and Kupffer cells; 2) acquisition, maintenance in culture, cryopreservation and distribution to users of cell lines; 3) preparation of hybridomas producing antibodies to user-provided antigens; 4) optimization of conditions for transfecting cell lines or primary cultures; 5) preparation/distribution of culture medium; 6) testing of cell lines for mycoplasma contamination; 7) provision of space, equipment and sterile supplies for cell culture work; and 8) provision of training, assistance and consultation in cell culture techniques. During the last three years, use of the facility has grown substantially, from three users at the time of the original grant application, to 18 investigators who have used the facility to date. Fourteen of the above eighteen investigators are Center members and these individuals account for 99% of the total usage of the CCFC. Future plans include the establishment of the following services, which are currently under development: 1) isolation and primary culture of human hepatocytes; 2) preparation of liver slices for short term studies; and 3) isolation of cells representing different stages of the cell cycle by centrifugal elutriation. The CCFC will also improve its ability to produce hybridomas and will continue to work with investigators to develop new cell isolation techniques.

细胞培养设施核心(CCFC)被安置在两个 化学毒理研究所的实验室，里面有所有 细胞培养工作所需的设备。另一个实验室 在韦恩已经建立了培养细胞的病毒转染法 州立大学医学院，但尚不清楚这在多大程度上 后者的实验室由CCFC控制。托马斯·科卡雷克博士是核心 领导人，并要求他的工资的20%为这一角色。他得到了 两名全职技术人员。该设施提供以下服务： 1)动物肝细胞和枯否细胞原代培养的制备； 2)获取、在培养中维护、超低温保存和分发给 细胞系使用者；3)产生抗体的杂交瘤的制备 用户提供的抗原；4)细胞转染条件的优化 品系或原代培养；5)培养基配制/分配； 6)检测细胞系的支原体污染；7)提供空间， 用于细胞培养工作的设备和无菌用品；以及8)提供 细胞培养技术方面的培训、帮助和咨询。 在过去三年中，该设施的使用量大幅增长，从 最初拨款申请时的三名用户，给18名调查人员 到目前为止已经使用过该设施的人。以上十八个中的十四个 调查人员是中心成员，这些个人占中心成员的99% CCFC的总使用量。未来的计划包括建立 目前正在开发的以下服务：1)隔离和 人肝细胞原代培养；2)肝切片制备 学期研究；以及3)分离代表不同阶段的细胞 用离心法洗脱细胞周期。CCFC还将提高其能力 生产杂交瘤，并将继续与调查人员合作开发 新的细胞分离技术。