歡迎您，來到青旗（上海）生物技術(shù)發(fā)展有限公司！
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簡(jiǎn)要描述：青旗（上海）生物技術(shù)發(fā)展有限公司，總部位于上海浦東新區(qū)，依托本地高校資源，逐步發(fā)展成為以生物技術(shù)為主的研發(fā)、生產(chǎn)、培訓(xùn)為一體的綜合化產(chǎn)業(yè)平臺(tái)，在標(biāo)準(zhǔn)化細(xì)胞庫建立及細(xì)胞藥物前端模型方面成果顯著。公司生產(chǎn)經(jīng)營(yíng)原代細(xì)胞、細(xì)胞系、ELISA試劑盒、感受態(tài)細(xì)胞和HPLC檢測(cè)等科研產(chǎn)品與服務(wù)。我們秉承對(duì)用戶負(fù)責(zé)的態(tài)度，以對(duì)科研的高度嚴(yán)謹(jǐn)，以嚴(yán)格的質(zhì)量控制，為廣大生物醫(yī)學(xué)科研用戶提供更優(yōu)質(zhì)的服務(wù)！
更新時(shí)間：2021-05-21
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| 品牌 | 其他品牌 | 貨號(hào) | BFN60800665 |
|---|---|---|---|
| 規(guī)格 | T25培養(yǎng)瓶x1 1.5ml凍存管x2 | 供貨周期 | 現(xiàn)貨 |
| 主要用途 | 僅供科研 | 應(yīng)用領(lǐng)域 | 醫(yī)療衛(wèi)生,生物產(chǎn)業(yè) |
細(xì)胞名稱 | 人肺癌細(xì)胞A549 |
| |
貨物編碼 | BFN60800665 | ||
產(chǎn)品規(guī)格 | T25培養(yǎng)瓶x1 | 1.5ml凍存管x2 | |
細(xì)胞數(shù)量 | 1x10^6 | 1x10^6 | |
保存溫度 | 37℃ | -198℃ | |
運(yùn)輸方式 | 常溫保溫運(yùn)輸 | 干冰運(yùn)輸 | |
安全等級(jí) | 1 | ||
用途限制 | 僅供科研用途 1類 | ||
培養(yǎng)體系 | DMEM高糖培養(yǎng)基（Hyclone）+10%胎牛血清（Gibco）+1%雙抗（Hyclone） | ||
培養(yǎng)溫度 | 37℃ | 二氧化碳濃度 | 5% |
簡(jiǎn)介 | 人肺癌細(xì)胞A549細(xì)胞系是1972年由GiardDJ通過肺癌組織移植培養(yǎng)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(xiàn) | 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(yàn)收細(xì)胞注意事項(xiàng)
1、收到人肺癌細(xì)胞A549細(xì)胞，請(qǐng)查看瓶子是否有破裂，培養(yǎng)基是否漏出，是否渾濁，如有請(qǐng)盡快聯(lián)系。
2、收到人肺癌細(xì)胞A549細(xì)胞，如包裝完好，請(qǐng)?jiān)陲@微鏡下觀察細(xì)胞。,由于運(yùn)輸過程中的問題，細(xì)胞培養(yǎng)瓶中的貼壁細(xì)胞有可能從瓶壁中脫落下來，顯微鏡下觀察會(huì)出現(xiàn)細(xì)胞懸浮的情況，出現(xiàn)此狀態(tài)時(shí)，請(qǐng)不要打開細(xì)胞培養(yǎng)瓶，應(yīng)立即將培養(yǎng)瓶置于細(xì)胞培養(yǎng)箱里靜止 3-5 小時(shí)左右，讓細(xì)胞先穩(wěn)定下，再于顯微鏡下觀察，此時(shí)多數(shù)細(xì)胞會(huì)重新貼附于瓶壁。如細(xì)胞仍不能貼壁，請(qǐng)用臺(tái)盼藍(lán)染色法鑒定細(xì)胞活力，如臺(tái)盼藍(lán)染色證實(shí)細(xì)胞活力正常請(qǐng)按懸浮細(xì)胞的方法處理。
3、收到人肺癌細(xì)胞A549細(xì)胞后，請(qǐng)鏡下觀察細(xì)胞，用恰當(dāng)方式處理細(xì)胞。若懸浮的細(xì)胞較多，請(qǐng)離心收集細(xì)胞，接種到一個(gè)新的培養(yǎng)瓶中。棄掉原液，使用新鮮配制的培養(yǎng)基，使用進(jìn)口胎牛血清。剛接到細(xì)胞，若細(xì)胞不多時(shí) 血清濃度可以加到 15％去培養(yǎng)。若細(xì)胞迏到 80%左右 ，血清濃度還是在 10％。
4、收到人肺癌細(xì)胞A549細(xì)胞時(shí)如無異常情況 ，請(qǐng)?jiān)陲@微鏡下觀察細(xì)胞密度，如為貼壁細(xì)胞，未超過80%匯合度時(shí)，將培養(yǎng)瓶中培養(yǎng)基吸出，留下 5-10ML 培養(yǎng)基繼續(xù)培養(yǎng)：超過 80%匯合度時(shí)，請(qǐng)按細(xì)胞培養(yǎng)條件傳代培養(yǎng)。如為懸浮細(xì)胞，吸出培養(yǎng)液，1000 轉(zhuǎn)/分鐘離心 3 分鐘，吸出上清，管底細(xì)胞用新鮮培養(yǎng)基懸浮細(xì)胞后移回培養(yǎng)瓶。
5、將培養(yǎng)瓶置于 37℃培養(yǎng)箱中培養(yǎng)，蓋子微微擰松。吸出的培養(yǎng)基可以保存在滅菌過的瓶子里，存放于 4℃冰箱，以備不時(shí)之需。
6、24 小時(shí)后，人肺癌細(xì)胞A549細(xì)胞形態(tài)已恢復(fù)并貼滿瓶壁，即可傳代。（貼壁細(xì)胞）將培養(yǎng)瓶里的培養(yǎng)基倒去，加 3-5ml（以能覆蓋細(xì)胞生長(zhǎng)面為準(zhǔn)）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(shí)間以具體細(xì)胞為準(zhǔn)，一般 1-3 分鐘，不超過 5 分鐘。可以放入37℃培養(yǎng)箱消化。輕輕晃動(dòng)瓶壁，見細(xì)胞脫落下來，加入 3-5ml 培養(yǎng)基終止消化。用移液管輕輕吹打瓶壁上的細(xì)胞，使之*脫落，然后將溶液吸入離心管內(nèi)離心，1000rpm/5min。棄上清，視細(xì)胞數(shù)量決定分瓶數(shù)，一般一傳二，如細(xì)胞量多可一傳三，有些細(xì)胞不易傳得過稀，有些生長(zhǎng)較快的細(xì)胞則可以多傳幾瓶，以具體細(xì)胞和經(jīng)驗(yàn)為準(zhǔn)。（懸浮細(xì)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(xì)胞 ，懸浮細(xì)胞。嚴(yán)格無菌操作。換液時(shí)，換新的細(xì)胞培養(yǎng)瓶和換新鮮的培養(yǎng)液，37℃，5%CO2 培養(yǎng)。
特別提醒： 原瓶中培養(yǎng)基不宜繼續(xù)使用，請(qǐng)更換新鮮培養(yǎng)基培養(yǎng)。
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