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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(kù)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) | BFN60800668 |
|---|---|---|---|
| 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 | 人非小細(xì)胞肺癌細(xì)胞HCC827 |
| |
貨物編碼 | BFN60800668 | ||
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 | 人非小細(xì)胞肺癌細(xì)胞HCC827 細(xì)胞建于1994年三月。這株肺腺癌在EGFR酪氨酸激酶區(qū)域有一個(gè)獲得性突變(E746-A750缺失)?；颊?/strong>在25歲到26歲時(shí)每個(gè)月抽1包煙。在診斷前12年不再抽煙。 | ||
注釋 | Part of: Cancer Cell Line Encyclopedia (CCLE) project. Part of: COSMIC cell lines project. Part of: MD Anderson Cell Lines Project. Doubling time: 71 hours (PubMed=25984343); 28 hours (ATCC); ~50-60 hours (DSMZ). Microsatellite instability: Stable (MSS) (Sanger). Omics: Deep exome analysis. Omics: Deep proteome analysis. Omics: Deep RNAseq analysis. Omics: DNA methylation analysis. Omics: Protein expression by reverse-phase protein arrays. Omics: shRNA library screening. Omics: SNP array analysis. Omics: Transcriptome analysis. | ||
STR信息 | Amelogenin:X；CSF1PO:11；D13S317:9；D16S539:12；D18S51:13；D19S433:14；D21S11:31；D2S1338:17,24；D3S1358:17；D5S818:12；D7S820:11,12；D8S1179:12；FGA:22,24；TH01:6；TPOX:8；vWA:18 | ||
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PubMed=22961666; DOI=10.1158/2159-8290.CD-12-0112 Byers L.A., Wang J., Nilsson M.B., Fujimoto J., Saintigny P., Yordy J., Giri U., Peyton M., Fan Y.H., Diao L., Masrorpour F., Shen L., Liu W., Duchemann B., Tumula P., Bhardwaj V., Welsh J., Weber S., Glisson B.S., Kalhor N., Wistuba I.I., Girard L., Lippman S.M., Mills G.B., Coombes K.R., Weinstein J.N., Minna J.D., Heymach J.V. Proteomic profiling identifies dysregulated pathways in small cell lung cancer and novel therapeutic targets including PARP1. Cancer Discov. 2:798-811(2012)
PubMed=23733853; DOI=10.1101/gr.152322.112 Jia P., Jin H., Meador C.B., Xia J., Ohashi K., Liu L., Pirazzoli V., Dahlman K.B., Politi K., Michor F., Zhao Z., Pao W. Next-generation sequencing of paired tyrosine kinase inhibitor-sensitive and -resistant EGFR mutant lung cancer cell lines identifies spectrum of DNA changes associated with drug resistance. Genome Res. 23:1434-1445(2013)
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PubMed=25984343; DOI=10.1038/sdata.2014.35 Cowley G.S., Weir B.A., Vazquez F., Tamayo P., Scott J.A., Rusin S., East-Seletsky A., Ali L.D., Gerath W.F.J., Pantel S.E., Lizotte P.H., Jiang G., Hsiao J., Tsherniak A., Dwinell E., Aoyama S., Okamoto M., Harrington W., Gelfand E., Green T.M., Tomko M.J., Gopal S., Wong T.C., Li H., Howell S., Stransky N., Liefeld T., Jang D., Bistline J., Hill Meyers B., Armstrong S.A., Anderson K.C., Stegmaier K., Reich M., Pellman D., Boehm J.S., Mesirov J.P., Golub T.R., Root D.E., Hahn W.C. Parallel genome-scale loss of function screens in 216 cancer cell lines for the identification of context-specific genetic dependencies. Sci. Data 1:140035-140035(2014)
PubMed=25485619; DOI=10.1038/nbt.3080 Klijn C., Durinck S., Stawiski E.W., Haverty P.M., Jiang Z., Liu H., Degenhardt J., Mayba O., Gnad F., Liu J., Pau G., Reeder J., Cao Y., Mukhyala K., Selvaraj S.K., Yu M., Zynda G.J., Brauer M.J., Wu T.D., Gentleman R.C., Manning G., Yauch R.L., Bourgon R., Stokoe D., Modrusan Z., Neve R.M., de Sauvage F.J., Settleman J., Seshagiri S., Zhang Z. A comprehensive transcriptional portrait of human cancer cell lines. Nat. Biotechnol. 33:306-312(2015)
PubMed=26361996; DOI=10.1016/j.jprot.2015.09.003 Grundner-Culemann K., Dybowski J.N., Klammer M., Tebbe A., Schaab C., Daub H. Comparative proteome analysis across non-small cell lung cancer cell lines. J. Proteomics 130:1-10(2016)
PubMed=27397505; DOI=10.1016/j.cell.2016.06.017 Iorio F., Knijnenburg T.A., Vis D.J., Bignell G.R., Menden M.P., Schubert M., Aben N., Goncalves E., Barthorpe S., Lightfoot H., Cokelaer T., Greninger P., van Dyk E., Chang H., de Silva H., Heyn H., Deng X., Egan R.K., Liu Q., Mironenko T., Mitropoulos X., Richardson L., Wang J., Zhang T., Moran S., Sayols S., Soleimani M., Tamborero D., Lopez-Bigas N., Ross-Macdonald P., Esteller M., Gray N.S., Haber D.A., Stratton M.R., Benes C.H., Wessels L.F.A., Saez-Rodriguez J., McDermott U., Garnett M.J. A landscape of pharmacogenomic interactions in cancer. Cell 166:740-754(2016)
PubMed=28196595; DOI=10.1016/j.ccell.2017.01.005 Li J., Zhao W., Akbani R., Liu W., Ju Z., Ling S., Vellano C.P., Roebuck P., Yu Q., Eterovic A.K., Byers L.A., Davies M.A., Deng W., Gopal Y.N.V., Chen G., von Euw E.M., Slamon D.J., Conklin D., Heymach J.V., Gazdar A.F., Minna J.D., Myers J.N., Lu Y., Mills G.B., Liang H. Characterization of human cancer cell lines by reverse-phase protein arrays. Cancer Cell 31:225-239(2017)
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驗(yàn)收細(xì)胞注意事項(xiàng)
1、收到人非小細(xì)胞肺癌細(xì)胞HCC827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、收到人非小細(xì)胞肺癌細(xì)胞HCC827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(guò)程中的問(wèn)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(lái)，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(kāi)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、收到人非小細(xì)胞肺癌細(xì)胞HCC827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、收到人非小細(xì)胞肺癌細(xì)胞HCC827細(xì)胞時(shí)如無(wú)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(guò)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(guò) 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(guò)的瓶子里，存放于 4℃冰箱，以備不時(shí)之需。
6、24 小時(shí)后，人非小細(xì)胞肺癌細(xì)胞HCC827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(guò) 5 分鐘?？梢苑?/span>入37℃培養(yǎng)箱消化。輕輕晃動(dòng)瓶壁，見(jiàn)細(xì)胞脫落下來(lái)，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(guò)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(wú)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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