| 品牌 | 其他品牌 | 貨號 | BFN60800682 |
|---|---|---|---|
| 規(guī)格 | T25培養(yǎng)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(yī)療衛(wèi)生,生物產業(yè) |
細胞名稱 | 人肝癌細胞SNU-475 |
| |
貨物編碼 | BFN60800682 | ||
產品規(guī)格 | T25培養(yǎng)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 3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% |
簡介 | 人肝癌細胞SNU-475取自亞洲人，男性。人肝癌細胞SNU-475由青旗（上海）生物技術發(fā)展有限公司于2018年引種自ATCC(CRL-2236)。 | ||
注釋 | Part of: Cancer Cell Line Encyclopedia (CCLE) project. Part of: COSMIC cell lines project. Part of: MD Anderson Cell Lines Project. Part of: Seoul National University (SNU) cell line collection. Characteristics: Has lost chromosome Y. Doubling time: 66 hours (PubMed=7543080). Microsatellite instability: Stable (MSS) (Sanger). Transformant: NCBI_TaxID; 10407; Hepatitis B virus (HBV). Omics: Array-based CGH. Omics: Deep exome analysis. Omics: Deep RNAseq analysis. Omics: DNA methylation analysis. Omics: Protein expression by reverse-phase protein arrays. Omics: SNP array analysis. Omics: Transcriptome analysis. | ||
STR信息 | Amelogenin: X,Y CSF1PO: 11,12 D13S317: 8,11 D16S539: 12 D5S818: 10,13 D7S820: 7,12 TH01: 7,9 TPOX: 8,9 vWA: 14 | ||
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PubMed=19956504; DOI=10.4143/crt.2005.37.1.1 Ku J.-L., Park J.-G. Biology of SNU cell lines. Cancer Res. Treat. 37:1-19(2005)
PubMed=16616112; DOI=10.1016/j.cancergencyto.2005.08.022 Park S.-J., Jeong S.-Y., Kim H.J. Y chromosome loss and other genomic alterations in hepatocellular carcinoma cell lines analyzed by CGH and CGH array. Cancer Genet. Cytogenet. 166:56-64(2006)
PubMed=20164919; DOI=10.1038/nature08768 Bignell G.R., Greenman C.D., Davies H., Butler A.P., Edkins S., Andrews J.M., Buck G., Chen L., Beare D., Latimer C., Widaa S., Hinton J., Fahey C., Fu B., Swamy S., Dalgliesh G.L., Teh B.T., Deloukas P., Yang F., Campbell P.J., Futreal P.A., Stratton M.R. Signatures of mutation and selection in the cancer genome. Nature 463:893-898(2010)
PubMed=20215515; DOI=10.1158/0008-5472.CAN-09-3458 Rothenberg S.M., Mohapatra G., Rivera M.N., Winokur D., Greninger P., Nitta M., Sadow P.M., Sooriyakumar G., Brannigan B.W., Ulman M.J., Perera R.M., Wang R., Tam A., Ma X.-J., Erlander M., Sgroi D.C., Rocco J.W., Lingen M.W., Cohen E.E.W., Louis D.N., Settleman J., Haber D.A. A genome-wide screen for microdeletions reveals disruption of polarity complex genes in diverse human cancers. Cancer Res. 70:2158-2164(2010)
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=23505090; DOI=10.1002/hep.26402 Wang K., Lim H.Y., Shi S., Lee J., Deng S., Xie T., Zhu Z., Wang Y., Pocalyko D., Yang W.J., Rejto P.A., Mao M., Park C.-K., Xu J. Genomic landscape of copy number aberrations enables the identification of oncogenic drivers in hepatocellular carcinoma. Hepatology 58:706-717(2013)
PubMed=23887712; DOI=10.1038/ncomms3218 Nault J.-C., Mallet M., Pilati C., Calderaro J., Bioulac-Sage P., Laurent C., Laurent A., Cherqui D., Balabaud C., Zucman-Rossi J. High frequency of telomerase reverse-transcriptase promoter somatic mutations in hepatocellular carcinoma and preneoplastic lesions. Nat. Commun. 4:2218-2218(2013)
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=25574106; DOI=10.3748/wjg.v21.i1.311 Cevik D., Yildiz G., Ozturk M. Common telomerase reverse transcriptase promoter mutations in hepatocellular carcinomas from different geographical locations. World J. Gastroenterol. 21:311-317(2015)
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=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、收到人肝癌細胞SNU-475細胞，請查看瓶子是否有破裂，培養(yǎng)基是否漏出，是否渾濁，如有請盡快聯系。
2、收到人肝癌細胞SNU-475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(yǎng)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(tài)時，請不要打開細胞培養(yǎng)瓶，應立即將培養(yǎng)瓶置于細胞培養(yǎng)箱里靜止 3-5 小時左右，讓細胞先穩(wěn)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、收到人肝癌細胞SNU-475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(yǎng)瓶中。棄掉原液，使用新鮮配制的培養(yǎng)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(yǎng)。若細胞迏到 80%左右 ，血清濃度還是在 10％。
4、收到人肝癌細胞SNU-475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(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，請按細胞培養(yǎng)條件傳代培養(yǎng)。如為懸浮細胞，吸出培養(yǎng)液，1000 轉/分鐘離心 3 分鐘，吸出上清，管底細胞用新鮮培養(yǎng)基懸浮細胞后移回培養(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，以備不時之需。
6、24 小時后，人肝癌細胞SNU-475細胞形態(tài)已恢復并貼滿瓶壁，即可傳代。（貼壁細胞）將培養(yǎng)瓶里的培養(yǎng)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(yǎng)箱消化。輕輕晃動瓶壁，見細胞脫落下來，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(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，請更換新鮮培養(yǎng)基培養(yǎng)。
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