| 品牌 | 其他品牌 | 货号 | BFN60800665 |
|---|---|---|---|
| 规格 | T25培养瓶x1 1.5ml冻存管x2 | 供货周期 | 现货 |
| 主要用途 | 仅供科研 | 应用领域 | 医疗卫生,生物产业 |
细胞名称 | 人肺癌细胞A549 |
| |
货物编码 | BFN60800665 | ||
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 | 人肺癌细胞A549细胞系是1972年由GiardDJ通过肺癌组织移植培养建系的，源自一位58岁的白人男性。A549能通过胞苷二磷脂酰胆碱途径合成富含不饱和脂肪酸的卵磷脂；角蛋白阳性。 | ||
注释 | Part of: Cancer Cell Line Encyclopedia (CCLE) project. Part of: COSMIC cell lines project. Part of: ENCODE project common cell types; tier 2. Part of: JFCR39 cancer cell line panel. Part of: KuDOS 95 cell line panel. Part of: MD Anderson Cell Lines Project. Part of: Naval Biosciences Laboratory (NBL) collection (transferred to ATCC in 1982). Part of: NCI-60 cancer cell line panel. Part of: NCI-7 clinical proteomics reference material cell line panel. Doubling time: 18 hours (in RPMI 1640 + 10% FBS), 37 hours (in ACL-3), 36 hours (in ACL-3+BSA) (PubMed=3940644); 27.0 hours (PubMed=8286010); 22 hours (PubMed=25984343); 27 hours (from cell counting), 27 hours (from absorbance) (DOI=10.5897/IJBMBR2013.0154); 22.9 hours (NCI-DTP); ~28 hours (CLS); ~40 hours (DSMZ). Microsatellite instability: Stable (MSS) (PubMed=12661003; Sanger). Omics: Acetylation analysis by proteomics. Omics: Array-based CGH. Omics: CNV analysis. Omics: Deep antibody staining analysis. Omics: Deep exome analysis. Omics: Deep phosphoproteome analysis. Omics: Deep membrane proteome analysis. Omics: Deep proteome analysis. Omics: Deep RNAseq analysis. Omics: DNA methylation analysis. Omics: Fluorescence phenotype profiling. Omics: H3K4me3 ChIP-seq epigenome analysis. Omics: H3K9ac ChIP-seq epigenome analysis. Omics: lncRNA expression profiling. Omics: Metabolome analysis. Omics: Protein expression by reverse-phase protein arrays. Omics: Proteome analysis by 2D-DE/MS. Omics: shRNA library screening. Omics: SNP array analysis. Omics: Transcriptome analysis. Omics: Virome analysis using proteomics. Misspelling: A594; In PubMed=18227028. Misspelling: A59; In PubMed=16354588. | ||
STR信息 | Amelogenin：X，Y；CSF1PO：10，12；D13S317：11；D16S539：11，12；D18S51：14，17；D19S433：13；D21S11：29；D2S1338：24；D3S1358：16；D5S818：11；D7S820：8，11；D8S1179：13，14；FGA：23；TH01：8，9.3；TPOX：8，11；vWA：14； | ||
参考文献 | PubMed=4357758; DOI=10.1093/jnci/51.5.1417 Giard D.J., Aaronson S.A., Todaro G.J., Arnstein P., Kersey J.H., Dosik H., Parks W.P. In vitro c*tion of human tumors: establishment of cell lines derived from a series of solid tumors. J. Natl. Cancer Inst. 51:1417-1423(1973)
PubMed=175022; DOI=10.1002/ijc.2910170110 Lieber M.M., Smith B.T., Szakal A., Nelson-Rees W.A., Todaro G.J. A continuous tumor-cell line from a human lung carcinoma with properties of type II alveolar epithelial cells. Int. J. Cancer 17:62-70(1976)
PubMed=327080; DOI=10.1093/jnci/59.1.221 Fogh J., Fogh J.M., Orfeo T. One hundred and twenty-seven cultured human tumor cell lines producing tumors in nude mice. J. Natl. Cancer Inst. 59:221-226(1977)
PubMed=833871; DOI=10.1093/jnci/58.2.209 Fogh J., Wright W.C., Loveless J.D. Absence of HeLa cell contamination in 169 cell lines derived from human tumors. J. Natl. Cancer Inst. 58:209-214(1977)
PubMed=842942; DOI=10.1164/arrd.1977.115.2.285 Smith B.T. Cell line A549: a model system for the study of alveolar type II cell function. Am. Rev. Respir. Dis. 115:285-293(1977)
PubMed=924690; DOI=10.1002/ijc.2910200505 Kerbel R.S., Pross H.F., Leibovitz A. Analysis of established human carcinoma cell lines for lymphoreticular-associated membrane receptors. Int. J. Cancer 20:673-679(1977)
PubMed=22282976; DOI=10.1093/carcin/1.1.21 Day R.S. III, Ziolkowski C.H.J., Scudiero D.A., Meyer S.A., Mattern M.R. Human tumor cell strains defective in the repair of alkylation damage. Carcinogenesis 1:21-32(1980)
PubMed=6935474; DOI=10.1093/jnci/66.2.239 Wright W.C., Daniels W.P., Fogh J. Distinction of seventy-one cultured human tumor cell lines by polymorphic enzyme analysis. J. Natl. Cancer Inst. 66:239-247(1981)
PubMed=7459858 Rousset M., Zweibaum A., Fogh J. Presence of glycogen and growth-related variations in 58 cultured human tumor cell lines of various tissue origins. Cancer Res. 41:1165-1170(1981)
PubMed=7065527; DOI=10.1164/arrd.1982.125.2.222 Hay R.J., Williams C.D., Macy M.L., Lavappa K.S. Cultured cell lines for research on pulmonary physiology available through the American Type Culture Collection. Am. Rev. Respir. Dis. 125:222-232(1982)
PubMed=6148444; DOI=10.1093/jnci/73.4.801 Morstyn G., Russo A., Carney D.N., Karawya E., Wilson S.H., Mitchell J.B. Heterogeneity in the radiation survival curves and biochemical properties of human lung cancer cell lines. J. Natl. Cancer Inst. 73:801-807(1984)
PubMed=6500159; DOI=10.1159/000163283 Gershwin M.E., Lentz D., Owens R.B. Relationship between karyotype of tissue culture lines and tumorigenicity in nude mice. Exp. Cell Biol. 52:361-370(1984)
PubMed=3518877; DOI=10.3109/07357908609038260 Fogh J. Human tumor lines for cancer research. Cancer Invest. 4:157-184(1986)
PubMed=3940644 Brower M., Carney D.N., Oie H.K., Gazdar A.F., Minna J.D. Growth of cell lines and clinical specimens of human non-small cell lung cancer in a serum-free defined medium. Cancer Res. 46:798-806(1986)
PubMed=3945555; DOI=10.1093/nar/14.2.843 Valenzuela D.M., Groffen J. Four human carcinoma cell lines with novel mutations in position 12 of c-K-ras oncogene. Nucleic Acids Res. 14:843-852(1986)
PubMed=3129183 Hubbard W.C., Alley M.C., McLemore T.L., Boyd M.R. Evidence for thromboxane biosynthesis in established cell lines derived from human lung adenocarcinomas. Cancer Res. 48:2674-2677(1988)
PubMed=3335022 Alley M.C., Scudiero D.A., Monks A., Hursey M.L., Czerwinski M.J., Fine D.L., Abbott B.J., Mayo J.G., Shoemaker R.H., Boyd M.R. Feasibility of drug screening with panels of human tumor cell lines using a microculture tetrazolium assay. Cancer Res. 48:589-601(1988)
PubMed=3413074; DOI=10.1073/pnas.85.16.6042 Pereira-Smith O.M., Smith J.R. Genetic analysis of indefinite division in human cells: identification of four complementation groups. Proc. Natl. Acad. Sci. U.S.A. 85:6042-6046(1988)
PubMed=2388294; DOI=10.1093/jnci/82.17.1420 McLemore T.L., Litterst C.L., Coudert B.P., Liu M.C., Hubbard W.C., Adelberg S., Czerwinski M., McMahon N.A., Eggleston J.C., Boyd M.R. Metabolic activation of 4-ipomeanol in human lung, primary pulmonary carcinomas, and established human pulmonary carcinoma cell lines. J. Natl. Cancer Inst. 82:1420-1426(1990)
PubMed=2041050; DOI=10.1093/jnci/83.11.757 Monks A., Scudiero D.A., Skehan P., Shoemaker R.H., Paull K., Vistica D.T., Hose C., Langley J., Cronise P., Vaigro-Wolff A., Gray-Goodrich M., Campbell H., Mayo J.G., Boyd M.R. Feasibility of a high-flux anticancer drug screen using a diverse panel of cultured human tumor cell lines. J. Natl. Cancer Inst. 83:757-766(1991)
PubMed=8286010 Kiura K., Watarai S., Shibayama T., Ohnoshi T., Kimura I., Yasuda T. Inhibitory effects of cholera toxin on in vitro growth of human lung cancer cell lines. Anticancer Drug Des. 8:417-428(1993)
PubMed=7736387; DOI=10.1002/1097-0142(19950515)75:10<2442::AID-CNCR2820751009>3.0.CO;2-Q Campling B.G., Sarda I.R., Baer K.A., Pang S.C., Baker H.M., Lofters W.S., Flynn T.G. Secretion of atrial natriuretic peptide and vasopressin by small cell lung cancer. Cancer 75:2442-2451(1995)
PubMed=8626706; DOI=10.1074/jbc.271.19.11477 Quinn K.A., Treston A.M., Unsworth E.J., Miller M.-J., Vos M., Grimley C., Battey J., Mulshine J.L., Cuttitta F. Insulin-like growth factor expression in human cancer cell lines. J. Biol. Chem. 271:11477-11483(1996)
PubMed=9023415; DOI=10.1006/cimm.1996.1062 Seki N., Hoshino T., Kikuchi M., Hayashi A., Itoh K. HLA-A locus-restricted and tumor-specific CTLs in tumor-infiltrating lymphocytes of patients with non-small cell lung cancer. Cell. Immunol. 175:101-110(1997)
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验收细胞注意事项
1、收到人肺癌细胞A549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、收到人肺癌细胞A549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、收到人肺癌细胞A549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、收到人肺癌细胞A549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，人肺癌细胞A549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*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。
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