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| 品牌 | 其他品牌 | 貨號 | BFN60800668 |
|---|---|---|---|
| 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 | 人非小細胞肺癌細胞HCC827 |
| |
貨物編碼 | BFN60800668 | ||
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 | 人非小細胞肺癌細胞HCC827 細胞建于1994年三月。這株肺腺癌在EGFR酪氨酸激酶區域有一個獲得性突變(E746-A750缺失)。患者在25歲到26歲時每個月抽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=22460905; DOI=10.1038/nature11003 Barretina J.G., Caponigro G., Stransky N., Venkatesan K., Margolin A.A., Kim S., Wilson C.J., Lehar J., Kryukov G.V., Sonkin D., Reddy A., Liu M., Murray L., Berger M.F., Monahan J.E., Morais P., Meltzer J., Korejwa A., Jane-Valbuena J., Mapa F.A., Thibault J., Bric-Furlong E., Raman P., Shipway A., Engels I.H., Cheng J., Yu G.K., Yu J., Aspesi P. Jr., de Silva M., Jagtap K., Jones M.D., Wang L., Hatton C., Palescandolo E., Gupta S., Mahan S., Sougnez C., Onofrio R.C., Liefeld T., MacConaill L.E., Winckler W., Reich M., Li N., Mesirov J.P., Gabriel S.B., Getz G., Ardlie K., Chan V., Myer V.E., Weber B.L., Porter J., Warmuth M., Finan P., Harris J.L., Meyerson M., Golub T.R., Morrissey M.P., Sellers W.R., Schlegel R., Garraway L.A. The Cancer Cell Line Encyclopedia enables predictive modelling of anticancer drug sensitivity. Nature 483:603-607(2012)
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)
PubMed=25960936; DOI=10.4161/21624011.2014.954893 Boegel S., Lower M., Bukur T., Sahin U., Castle J.C. A catalog of HLA type, HLA expression, and neo-epitope candidates in human cancer cell lines. OncoImmunology 3:E954893-E954893(2014)
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)
PubMed=29444439; DOI=10.1016/j.celrep.2018.01.051 Yuan T.L., Amzallag A., Bagni R., Yi M., Afghani S., Burgan W., Fer N., Strathern L.A., Powell K., Smith B., Waters A.M., Drubin D., Thomson T., Liao R., Greninger P., Stein G.T., Murchie E., Cortez E., Egan R.K., Procter L., Bess M., Cheng K.T., Lee C.-S., Lee L.C., Fellmann C., Stephens R., Luo J., Lowe S.W., Benes C.H., McCormick F. Differential effector engagement by oncogenic KRAS. Cell Rep. 22:1889-1902(2018)
PubMed=30894373; DOI=10.1158/0008-5472.CAN-18-2747 Dutil J., Chen Z., Monteiro A.N., Teer J.K., Eschrich S.A. An interactive resource to probe genetic diversity and estimated ancestry in cancer cell lines. Cancer Res. 79:1263-1273(2019)
PubMed=31068700; DOI=10.1038/s41586-019-1186-3 Ghandi M., Huang F.W., Jane-Valbuena J., Kryukov G.V., Lo C.C., McDonald E.R. III, Barretina J., Gelfand E.T., Bielski C.M., Li H., Hu K., Andreev-Drakhlin A.Y., Kim J., Hess J.M., Haas B.J., Aguet F., Weir B.A., Rothberg M.V., Paolella B.R., Lawrence M.S., Akbani R., Lu Y., Tiv H.L., Gokhale P.C., de Weck A., Mansour A.A., Oh C., Shih J., Hadi K., Rosen Y., Bistline J., Venkatesan K., Reddy A., Sonkin D., Liu M., Lehar J., Korn J.M., Porter D.A., Jones M.D., Golji J., Caponigro G., Taylor J.E., Dunning C.M., Creech A.L., Warren A.C., McFarland J.M., Zamanighomi M., Kauffmann A., Stransky N., Imielinski M., Maruvka Y.E., Cherniack A.D., Tsherniak A., Vazquez F., Jaffe J.D., Lane A.A., Weinstock D.M., Johannessen C.M., Morrissey M.P., Stegmeier F., Schlegel R., Hahn W.C., Getz G., Mills G.B., Boehm J.S., Golub T.R., Garraway L.A., Sellers W.R. Next-generation characterization of the Cancer Cell Line Encyclopedia. Nature 569:503-508(2019) | ||
驗收細胞注意事項
1、收到人非小細胞肺癌細胞HCC827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、收到人非小細胞肺癌細胞HCC827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、收到人非小細胞肺癌細胞HCC827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、收到人非小細胞肺癌細胞HCC827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，人非小細胞肺癌細胞HCC827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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