芒果实验室，293细胞背景
培养过细胞的小伙伴们，肯定听说过293细胞 (全名HEK293)，但至于293细胞家族的其他衍生细胞却未必熟悉。
293细胞的衍生细胞可分为数种，比如293T细胞，293A细胞，293E细胞，293FT细胞等等。有网友总结说，293A是用来包腺病毒的，293T是用来包慢病毒的。总体而言，293细胞容易转染，是实验室最常用的表达兴趣基因的细胞株。
HEK293细胞是原代人胚肾细胞转染5型腺病毒DNA（Ad 5 DNA）而得到的永生化细胞，表达稳转的Ad5 DNA的基因，不易贴壁，表达E1A蛋白。
The human embryonic kidney (HEK) 293 cell line was derived from human embryonic kidney cells grown in tissue culture. The cell line was initiated by the transformation and culturing of normal HEK cells with sheared adenovirus 5 DNA. A portion of human adenovirus 5, nucleotides (nts) 1-4344, is integrated into chromosome 19 of the HEK cells. Human adenovirus 5 causes a range of respiratory illnesses.
HEK293T细胞由HEK293细胞衍生而来，含有SV40 large T抗原，贴壁不牢。293T细胞表达 E1A蛋白，S40大T抗原，含有S40复制起始点与启动子区的质粒可以复制。293T细胞容易转染，用磷酸钙、PEI或脂质体转染，效率均可达90%以上。
The 293T cell line was created in the laboratory of Michele Calos by transfection of a sub-line of adenovirus-immortalized human embryonic kidney cells with a gene encoding the SV40 T-antigen and a neomycin resistance gene. The cell line is competent for replication of vectors carrying the SV40 origin of replication. The line also has favorable tissue culture, transfection, DNA replication, gene expression, and protein production properties. It gives high titers when used to produce many viral vectors such as retroviruses and lentiviruses.
HEK293A细胞是HEK293细胞的亚株， A是Adhere单词的开头字母，倾向于形成单层细胞（monolayer）。在进行病毒滴度测定的空斑试验（plaque assay)时，293A细胞是均匀的单层，没有细胞重叠和间隙。通常用于腺病毒包装。与293A细胞所对的是293S（suspension)细胞，缺少N-乙酰氨基葡萄糖基转移酶，悬浮培养。
The HEK293S GnTIˉ cell line was established by methanesulfonate mutagenesis followed by Ricin selection. HEK293S GnTIˉ cells do not have N-acetyl-glucosaminyl transferase I (GnTI) activity and therefore lack complex N-glycans.
HEK293E细胞时稳定表达EBV病毒蛋白EBNA1抗原的HEK293细胞，悬浮培养，多用来表达真核蛋白。
293E cells are human embryonic kidney (HEK) cells derived from the 293 cell line and used for propagation of plasmids and expression of recombinant proteins in mammalian cells. The cell line expresses the EBNA-1 protein for episomal replication of oriP-harboring plasmids. The cell line is adapted for growth in suspension and for use with the mammalian expression system.
293FT细胞能制造高滴度慢病毒（F是Fast growth的首字母）。
The 293FT Cell Line is a very suitable host for lentiviral production. The 293FT Cell Line is derived from the 293F Cell Line and stably expresses the SV40 large T antigen from the pCMVSPORT6TAg.neo plasmid. Expression of the SV40 large T antigen is controlled by the human cytomegalovirus (CMV) promoter and is high-level and constitutive. The 293-F Cell Line is a fast-growing variant of the 293 cell line, and was originally obtained from Robert Horlick at Pharmacopeia.
参考资料
- https://ximbio.com/reagent/154155/hek-293s-cell-line
- https://www.hek293.com/
- https://www.thermofisher.com/cn/zh/home/references/protocols/cell-culture/transfection-protocol/growth-and-maintenance-of-the-293ft-cell-line.html