PA317 cell PA317 细胞 NICR865 cell NICR865 细胞 3142C0001000000097 Base Medium+8%DMSO+20%FBS C17 Cell Library of China Center For Type Culture Collection Class A Congyi Zheng, Sanfu Qu, Cell Library of China Center For Type Culture Collection, Wuhan University, 430072, 027－68754712, Fax:027－68754833, cctcc@whu.edu.cn MEM-EBSS: Minimum Essential Medium (MEM Eagles with Earle＇s Balanced Salts) 10%FBS Mouse Embryonic Fibroblast; PA317 NICR865 Negative PA317 PA317 cell PA317 cells derived from TK-NIH/3T3 cells were constructed by co-transfection of the packaging structure DNA (pPAM3) in pBR322 and the herpes simplex virus thymidine kinase gene in pBR322. The introduction of retroviral vectors into these cells by infection or transfection results in the formation of amphotropic vector particles that can infect many mammalian cells. Viral particles generated from the cells have been successfully used to introduce genes into humans. The percentage of PA317 cells capable of packaging retroviral vectors decreases with passage, possibly due to the loss of transferred DNA used to construct this strain of cells. A brief (approximately 5 days) selection in media containing 0.03 mM hypoxanthine, 0.001 mM methotrexate and 0.02 mM thymidine can select the ability of retaining the packaging function. The screened cells should be cultured in HT medium for 4 days to dilute the residual methotrexate. Passage at 1: 3, and passage once in 2-3 days. STR Identification The cell line (strain) of the library for research purposes only, may not be used for other purposes without permission. Users may not transfer the library cell line (strain) to a third party. Users in the publication of research papers or results, should indicate the source of cell lines (strains). adherent growth fibroblast-like http://www.cellresource.cn/fdetail.aspx?id=865 http://www.cellresource.cn/pdf/20080108100951.pdf 1:3传代,2-3天传一代 A类 C17 MEM-EBSS: Minimum Essential Medium (MEM Eagles with Earle＇s Balanced Salts) 10%FBS PA317 细胞 PA317细胞株源自TK- NIH/3T3细胞，通过pBR322中的包装结构DNA(pPAM3)和pBR322中的单纯疱疹病毒胸苷激酶基因共同转染构建而成。通过感染或转染将逆转录病毒载体导入这些细胞，生成可以感染许多哺乳动物细胞的双嗜性载体颗粒。这株细胞生成的病毒颗粒已成功地用于将基因导入人体。可能因为转入DNA丢失，能够包装逆转录病毒载体的PA317细胞百分比随传代而减少。在含0.03 mM 次黄嘌呤, 0.001 mM 氨甲喋呤和0.02 mM 胸苷的培养基中短暂(大约5天)选择，可以选出保留了包装功 STR鉴定 http://www.cellresource.cn/pdf/20080108100951.pdf 中国典型培养物保藏中心细胞库 基础培养基+8%DMSO+20%FBS 小鼠胚胎成纤维细胞；PA317 成纤维细胞样 本库的细胞系(株)仅用于研究工作，未经许可不得用于其它目的。使用者不得将本库细胞系(株)转让给第三者。使用者在发表研究论文或结果时，应注明细胞系(株)的来源。 贴壁生长 郑从义，屈三甫 武汉大学中国典型培养物保藏中心，430072，027－68754712传真027－68754833，cctcc@whu.edu.cn 阴性