| 品牌 | 其他品牌 | 貨號 | BFN60800665 |
|---|---|---|---|
| 規格 | T25培養瓶x1 1.5ml凍存管x2 | 供貨周期 | 現貨 |
| 主要用途 | 僅供科研 | 應用領域 | 醫療衛生,生物產業 |
細胞名稱 | 人肺癌細胞A549 |
| |
貨物編碼 | BFN60800665 | ||
產品規格 | T25培養瓶x1 | 1.5ml凍存管x2 | |
細胞數量 | 1x10^6 | 1x10^6 | |
保存溫度 | 37℃ | -198℃ | |
運輸方式 | 常溫保溫運輸 | 干冰運輸 | |
安全等級 | 1 | ||
用途限制 | 僅供科研用途 1類 | ||
培養體系 | DMEM高糖培養基（Hyclone）+10%胎牛血清（Gibco）+1%雙抗（Hyclone） | ||
培養溫度 | 37℃ | 二氧化碳濃度 | 5% |
簡介 | 人肺癌細胞A549細胞系是1972年由GiardDJ通過肺癌組織移植培養建系的，源自一位58歲的白人男性。A549能通過胞苷二磷脂酰膽堿途徑合成富含不飽和脂肪酸的卵磷脂；角蛋白陽性。 | ||
注釋 | 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； | ||
參考文獻 | 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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驗收細胞注意事項
1、收到人肺癌細胞A549細胞，請查看瓶子是否有破裂，培養基是否漏出，是否渾濁，如有請盡快聯系。
2、收到人肺癌細胞A549細胞，如包裝完好，請在顯微鏡下觀察細胞。,由于運輸過程中的問題，細胞培養瓶中的貼壁細胞有可能從瓶壁中脫落下來，顯微鏡下觀察會出現細胞懸浮的情況，出現此狀態時，請不要打開細胞培養瓶，應立即將培養瓶置于細胞培養箱里靜止 3-5 小時左右，讓細胞先穩定下，再于顯微鏡下觀察，此時多數細胞會重新貼附于瓶壁。如細胞仍不能貼壁，請用臺盼藍染色法鑒定細胞活力，如臺盼藍染色證實細胞活力正常請按懸浮細胞的方法處理。
3、收到人肺癌細胞A549細胞后，請鏡下觀察細胞，用恰當方式處理細胞。若懸浮的細胞較多，請離心收集細胞，接種到一個新的培養瓶中。棄掉原液，使用新鮮配制的培養基，使用進口胎牛血清。剛接到細胞，若細胞不多時 血清濃度可以加到 15％去培養。若細胞迏到 80%左右 ，血清濃度還是在 10％。
4、收到人肺癌細胞A549細胞時如無異常情況 ，請在顯微鏡下觀察細胞密度，如為貼壁細胞，未超過80%匯合度時，將培養瓶中培養基吸出，留下 5-10ML 培養基繼續培養：超過 80%匯合度時，請按細胞培養條件傳代培養。如為懸浮細胞，吸出培養液，1000 轉/分鐘離心 3 分鐘，吸出上清，管底細胞用新鮮培養基懸浮細胞后移回培養瓶。
5、將培養瓶置于 37℃培養箱中培養，蓋子微微擰松。吸出的培養基可以保存在滅菌過的瓶子里，存放于 4℃冰箱，以備不時之需。
6、24 小時后，人肺癌細胞A549細胞形態已恢復并貼滿瓶壁，即可傳代。（貼壁細胞）將培養瓶里的培養基倒去，加 3-5ml（以能覆蓋細胞生長面為準）PBS 或 Hanks’液洗滌后棄去。加 0.5-1ml 0.25%含 EDTA 的胰酶消化，消化時間以具體細胞為準，一般 1-3 分鐘，不超過 5 分鐘。可以放入37℃培養箱消化。輕輕晃動瓶壁，見細胞脫落下來，加入 3-5ml 培養基終止消化。用移液管輕輕吹打瓶壁上的細胞，使之*脫落，然后將溶液吸入離心管內離心，1000rpm/5min。棄上清，視細胞數量決定分瓶數，一般一傳二，如細胞量多可一傳三，有些細胞不易傳得過稀，有些生長較快的細胞則可以多傳幾瓶，以具體細胞和經驗為準。（懸浮細胞）用移液管輕輕吹打瓶壁，直接將溶液吸入離心管離心即可。
7、貼壁細胞 ，懸浮細胞。嚴格無菌操作。換液時，換新的細胞培養瓶和換新鮮的培養液，37℃，5%CO2 培養。
特別提醒： 原瓶中培養基不宜繼續使用，請更換新鮮培養基培養。
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