Core--Mammalian Cell Resource

哺乳动物细胞资源(MCR)核心在计划项目中有两个主要目标和几个功能。它的两个主要职责是：(1)彻底和仔细地描述在实施计划项目的研究计划过程中开发的哺乳动物细胞系，以及项目负责人确定的需要更广泛表征的任何其他细胞系，以及(2)根据个别项目的要求从特定细胞系制备哺乳动物细胞提取物。此外，MCR核心将为所有项目提供服务，将细胞系作为冷冻保存的样本进行维护，并根据需要为项目负责人确定的实验目的提供它们。这些冷冻样本将来自具有表型特征的细胞群体，并被证明不含支原体。通过集中和维护所有项目中使用的细胞系的质量控制，将促进整个计划项目的生化和遗传实验结果的比较，结果的重复性将得到保证，个别研究人员将对项目间的比较充满信心。MCR核心的目标是：(1)根据生长特性、克隆存活/细胞毒性和突变频率反应(例如，UVC、MMC、补骨脂素+UVA、IR或其他所需的细胞)、耐药表型(例如8-AA、6-TG、HAT、ALASA、G418等)和错配修复状态/错配修复表型，彻底而仔细地鉴定将在计划项目中广泛使用的所有细胞系。这些具有特征的细胞系随后将在培养中扩展，并将根据个别项目的要求准备和编目大量冷冻保存的样本，以便分发给各个项目；以及(2)根据各个项目的需要，制备小的(2升)和大的(最多50升)哺乳动物细胞系的提取物，用于进行生化分析(例如修复、切割、凝胶迁移率变化)。

The Mammalian Cell Resource (MCR) Core has two major goals and several functions in the Program Project. Its two major responsibilities are: (1) To thoroughly and carefully characterize the mammalian cell lines developed in the course of implementing the research plans within the Program Project, as well as any other cell lines that require more extensive characterization as determined by the Project Leaders, and (2) To prepare mammalian cell extracts from specific cell lines as required by the individual Projects. In addition, the MCR Core will provide the service to all Projects of maintaining cell lines as cryopreserved samples and supplying them on demand for the experimental purposes determined by the Project Leaders. These frozen samples will be derived from cell populations characterized phenotypically and certified free of mycoplasma. By centralizing and maintaining quality control of the cell lines to be used in all of the Projects, comparisons of the results of biochemical and genetic experiments across the Program Project will be facilitated, reproducibility of results will be assured, and individual investigators will be confident of inter-project comparisons. The aims of the MCR Core are: (1) To thoroughly and carefully characterize all cell lines that will be extensively used in the Program Project with respect to growth characteristics, clonogenic survival/cytotoxicity and mutation frequency responses (e.g., UVC, MMC, psoralen + UVA, IR, or others as needed), drug resistance phenotypes (e.g., 8-AA, 6-TG, HAT, ALASA, G418, etc.), and mismatch repair status/mismatch repair phenotypes, as appropriate. These characterized cell lines will then be expanded in culture and numerous cryopreserved samples from exponentially growing populations will be prepared and catalogued for distribution to the individual Projects as requested; and (2) To prepare both small (2-liter) and large (up to 50-liter) extracts from mammalian cell lines for the purposes of conducting biochemical assays (e.g., repair, incision, electrophoretic mobility shifts) as required by the individual Projects.