SV40转化的非洲绿猴肾细胞COS-7

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BLUEFBIO™ Product Sheet

细胞名称	SV40转化的非洲绿猴肾细胞COS-7
货物编码	BFN608006384
产品规格	T25培养瓶x1	1.5ml冻存管x2
细胞数量	1x10^6	1x10^6
保存温度	37℃	-198℃
运输方式	常温保温运输	干冰运输
安全等级	1
用途限制	仅供科研用途 1类

培养体系	DMEM高糖培养基（Hyclone）+10%胎牛血清（Gibco）+1%双抗（Hyclone）
培养温度	37℃	二氧化碳浓度	5%
简介	SV40转化的非洲绿猴肾细胞COS-7细胞株源自CV-1细胞株，经转染起始点缺失的SV40病毒突变体得到；编码表达野生型T抗原，所以该细胞适合作为需要SV40T抗原表达的载体的转染宿主。该细胞表达T抗原，允许SV40病毒的溶解性生长，支持40℃时温度敏感性A209病毒的复制，支持起始区域缺陷的SV40突变体的复制。因含有SV40病毒的DNA序列，该细胞需要在2级生物安全柜中操作。 SV40转化的非洲绿猴肾细胞COS-7细胞由青旗（上海）生物技术发展有限公司于2019年引种自ATCC(CRL-1651)
注释	Group: Non-human primate cell line. Doubling time: ~35-48 hours (DSMZ), ~2 days (lot 03052018), ~30 hours (lots 10052007 and 11222004), ~34 hours (lot 02292000) (JCRB). Transformant: NCBI_TaxID; 1891767; Simian virus 40 (SV40) [pSV6-1]. Misspelling: COS-2; In PubMed=14662750, PubMed=17074883 and PubMed=19379694, personal communication of Galan J.
STR信息	/
参考文献	Gluzman Y. SV40-transformed simian cells support the replication of early SV40 mutants. Cell 23: 175-182, 1981. PubMed: 6260373 Fernandez LM, Puett D. Lys583 in the third extracellular loop of the lutropinoriogonadotropin receptor is critical for signaling. J. Biol. Chem. 271: 925-930, 1996. PubMed: 8557706 Maestrini E, et al. A family of transmembrane proteins with homology to the MET-hepatocyte growth factor receptor. Proc. Natl. Acad. Sci. USA 93: 674-678, 1996. PubMed: 8570614 Campbell M, et al. The simian foamy virus type 1 transcriptional transactivator (Tas) binds and activates an enhancer element in the gag gene. J. Virol. 70: 6847-6855, 1996. PubMed: 8794326 Gonzalez Armas JC, et al. DNA immunization confers protection against murine cytomegalovirus infection. J. Virol. 70: 7921-7928, 1996. PubMed: 8892915 Jang SI, et al. Activator protein 1 activity is involved in the regulation of the cell type-specific expression from the proximal promoter of the human profilaggrin gene. J. Biol. Chem. 271: 24105-24114, 1996. PubMed: 8798649 Dittrich E, et al. A di-leucine motif and an upstream serine in the interleukin-6 (IL-6) signal transducer gp130 mediate ligand-induced endocytosis and down-regulation of the IL-6 receptor. J. Biol. Chem. 271: 5487-5494, 1996. PubMed: 8621406 Lee JH, et al. The proximal promoter of the human transglutaminase 3 gene. J. Biol. Chem. 271: 4561-4568, 1996. PubMed: 8626812 Chen Y, et al. Demonstration of binding of dengue virus envelope protein to target cells. J. Virol. 70: 8765-8772, 1996. PubMed: 8971005 Russell DW, Miller AD. Foamy virus vectors. J. Virol. 70: 217-222, 1996. PubMed: 8523528 Wright DA, et al. Association of human fas (CD95) with a ubiquitin-conjugating enzyme (UBC-FAP). J. Biol. Chem. 49: 31037-31043, 1996. PubMed: 8940097 Zhang J, et al. Dynamin and beta-arrestin reveal distinct mechanisms for G protein-coupled receptor internalization. J. Biol. Chem. 271: 18302-18305, 1996. PubMed: 8702465 Ozcelebi F, et al. Phosphorylation of cholecystokinin receptors expressed on chinese hamster ovary cells. J. Biol. Chem. 271: 3750-3755, 1996. PubMed: 8631990 Gibson S, et al. Functional LCK is required for optimal CD28-mediated activation of the TEC family tyrosine kinase EMT/ITK. J. Biol. Chem. 271: 7079-7083, 1996. PubMed: 8636141 Shaul PW, et al. Acylation targets endothelial nitric-oxide synthase to plasmalemmal caveolae. J. Biol. Chem. 271: 6518-6522, 1996. PubMed: 8626455 Ladner RD, et al. Identification of a consensus cyclin-dependent kinase phosphorylation site unique to the nuclear form of human deoxyuridine triphosphate nucleotidohydrolase. J. Biol. Chem. 271: 7752-7757, 1996. PubMed: 8631817 Wu X, et al. Demonstration of a physical interaction between microsomal triglyceride transfer protein and apolipoprotein B during the assembly of ApoB-containing lipoproteins. J. Biol. Chem. 271: 10277-10281, 1996. PubMed: 8626595 Hawes BE, et al. Phosphatidylinositol 3-kinase is an early intermediate in the G beta gamma-mediated mitogen-activated protein kinase signaling pathway. J. Biol. Chem. 271: 12133-12136, 1996. PubMed: 8647803 Arai H, Charo IF. Differential regulation of G-protein-mediated signaling by chemokine receptors. J. Biol. Chem. 271: 21814-21819, 1996. PubMed: 8702980 Hsieh CM, et al. APEG-1, a novel gene preferentially expressed in aortic smooth muscle cells, is down-regulated by vascular injury. J. Biol. Chem. 271: 17354-17359, 1996. PubMed: 8663449 Holtmann MH, et al. Multiple extracellular loop domains contribute critical determinants for agonist binding and activation of the secretin receptor. J. Biol. Chem. 271: 14944-14949, 1996. PubMed: 8663161

验收细胞注意事项

1、收到SV40转化的非洲绿猴肾细胞COS-7细胞，请查看瓶子是否有破裂，培养基是否漏出，是否浑浊，如有请尽快联系。

2、收到SV40转化的非洲绿猴肾细胞COS-7细胞，如包装完好，请在显微镜下观察细胞。,由于运输过程中的问题，细胞培养瓶中的贴壁细胞有可能从瓶壁中脱落下来，显微镜下观察会出现细胞悬浮的情况，出现此状态时，请不要打开细胞培养瓶，应立即将培养瓶置于细胞培养箱里静止 3-5 小时左右，让细胞先稳定下，再于显微镜下观察，此时多数细胞会重新贴附于瓶壁。如细胞仍不能贴壁，请用台盼蓝染色法鉴定细胞活力，如台盼蓝染色证实细胞活力正常请按悬浮细胞的方法处理。

3、收到SV40转化的非洲绿猴肾细胞COS-7细胞后，请镜下观察细胞，用恰当方式处理细胞。若悬浮的细胞较多，请离心收集细胞，接种到一个新的培养瓶中。弃掉原液，使用新鲜配制的培养基，使用进口胎牛血清。刚接到细胞，若细胞不多时血清浓度可以加到 15％去培养。若细胞迏到 80%左右，血清浓度还是在 10％。

4、收到SV40转化的非洲绿猴肾细胞COS-7细胞时如无异常情况，请在显微镜下观察细胞密度，如为贴壁细胞，未超过80%汇合度时，将培养瓶中培养基吸出，留下 5-10ML 培养基继续培养：超过 80%汇合度时，请按细胞培养条件传代培养。如为悬浮细胞，吸出培养液，1000 转/分钟离心 3 分钟，吸出上清，管底细胞用新鲜培养基悬浮细胞后移回培养瓶。

5、将培养瓶置于 37℃培养箱中培养，盖子微微拧松。吸出的培养基可以保存在灭菌过的瓶子里，存放于 4℃冰箱，以备不时之需。

6、24 小时后，SV40转化的非洲绿猴肾细胞COS-7细胞形态已恢复并贴满瓶壁，即可传代。（贴壁细胞）将培养瓶里的培养基倒去，加 3-5ml（以能覆盖细胞生长面为准）PBS 或 Hanks’液洗涤后弃去。加 0.5-1ml 0.25%含 EDTA 的胰酶消化，消化时间以具体细胞为准，一般 1-3 分钟，不超过 5 分钟。可以放入37℃培养箱消化。轻轻晃动瓶壁，见细胞脱落下来，加入 3-5ml 培养基终止消化。用移液管轻轻吹打瓶壁上的细胞，使之完全脱落，然后将溶液吸入离心管内离心，1000rpm/5min。弃上清，视细胞数量决定分瓶数，一般一传二，如细胞量多可一传三，有些细胞不易传得过稀，有些生长较快的细胞则可以多传几瓶，以具体细胞和经验为准。（悬浮细胞）用移液管轻轻吹打瓶壁，直接将溶液吸入离心管离心即可。

7、贴壁细胞，悬浮细胞。严格无菌操作。换液时，换新的细胞培养瓶和换新鲜的培养液，37℃，5%CO2 培养。

特别提醒： 原瓶中培养基不宜继续使用，请更换新鲜培养基培养。