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簡要描述: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)、培訓為一體的綜合化產(chǎn)業(yè)平臺,在標準化細胞庫建立及細胞藥物前端模型方面成果顯著。公司生產(chǎn)經(jīng)營原代細胞、細胞系、ELISA試劑盒、感受態(tài)細胞和HPLC檢測等科研產(chǎn)品與服務(wù)。我們秉承對用戶負責的態(tài)度,以對科研的高度嚴謹,以嚴格的質(zhì)量控制,為廣大生物醫(yī)學科研用戶提供更優(yōu)質(zhì)的服務(wù)!
更新時間:2021-05-25
廠商性質(zhì):生產(chǎn)廠家
瀏覽次數(shù):334
品牌 | 其他品牌 | 貨號 | BFN60800646 |
---|---|---|---|
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) |
細胞名稱 | 人結(jié)腸癌細胞HT-29 | ||
貨物編碼 | BFN60800646 | ||
產(chǎn)品規(guī)格 | T25培養(yǎng)瓶x1 | 1.5ml凍存管x2 | |
細胞數(shù)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(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 | 人結(jié)腸癌細胞HT-29細胞是1964年由FoghJ用移植培養(yǎng)方法和含15%FBS的F12培養(yǎng)液從原發(fā)性腫瘤分離的。近來,已建株的培養(yǎng)細胞用含血清的McCoy's5a培養(yǎng)基培養(yǎng)。該細胞系在裸鼠中成瘤,也能在類固醇處理的地鼠中成瘤。該細胞可合成IgA、CEA、TGFβ結(jié)合蛋白和黏液素;表達尿激酶受體,但沒有檢測到血漿酶原活性;不表達CD4,但細胞表面表達半乳糖神經(jīng)酰胺(HIV的可能替代受體)。該細胞系癌基因c-myc、K-ras、H-ras、N-ras、Myb、sis、fos陽性;p53基因過表達。人結(jié)腸癌細胞HT-29細胞由青旗(上海)生物技術(shù)發(fā)展有限公司于2017年引種自ATCC(HTB-38)。 | ||
注釋 | Part of: AstraZeneca Colorectal cell line (AZCL) panel. Part of: Cancer Cell Line Encyclopedia (CCLE) project. Part of: COSMIC cell lines project. Part of: JFCR39 cancer cell line panel. Part of: KuDOS 95 cell line panel. Part of: MD Anderson Cell Lines Project. Part of: NCI-60 cancer cell line panel. From: Memorial Sloan Kettering Cancer Center; New York; USA. Registration: Memorial Sloan Kettering Cancer Center Office of Technology Development; SK 809. Characteristics: Hypertriploid karyotype. Doubling time: 24 hours (PubMed=25984343); 24 hours (PubMed=23546019); 19.5 hours (NCI-DTP); ~24 hours (CLS); ~40-60 hours (DSMZ); ~23 hours (PBCF). Microsatellite instability: Stable (MSS) (PubMed=9000147; PubMed=24042735; PubMed=25926053; PubMed=28683746; Sanger). Omics: Array-based CGH. Omics: CNV analysis. Omics: Deep exome analysis. Omics: Deep phosphoproteome analysis. Omics: Deep proteome analysis. Omics: Deep quantitative phosphoproteome analysis. Omics: Deep quantitative proteome analysis. Omics: Deep RNAseq analysis. Omics: DNA methylation analysis. Omics: Fluorescence phenotype profiling. Omics: lncRNA expression profiling. Omics: Metabolome analysis. Omics: miRNA expression profiling. Omics: N-glycan profiling. Omics: Protein expression by reverse-phase protein arrays. Omics: shRNA library screening. Omics: SNP array analysis. Omics: Transcriptome analysis. Caution: Indicated in PubMed=11921276 as originating from an adenocarcinoma of the rectosigmoid part of the intestine, but is only indicated in the oldest publication (DOI=10.1007/978-1-4757-1647-4_5) as a colon adenocarcinoma. Misspelling: HCT29; In GEO GSM206503. | ||
STR信息 | Amelogenin:X;CSF1PO:11,12;D13S317:11,12;D16S539:11,12;D18S51:13;D19S433:14;D21S11:29,30;D2S1338:19,23;D3S1358:15,16.3;D5S818:11,12;D7S820:10;D8S1179:10,16;FGA:20,22;TH01:6,9;TPOX:8,9;vWA:17,19; | ||
參考文獻 | PubMed=27377824; DOI=10.1038/sdata.2016.52 Mestdagh P., Lefever S., Volders P.-J., Derveaux S., Hellemans J., Vandesompele J. Long non-coding RNA expression profiling in the NCI60 cancer cell line panel using high-throughput RT-qPCR. Sci. Data 3:160052-160052(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=27807467; DOI=10.1186/s13100-016-0078-4 Zampella J.G., Rodic N., Yang W.R., Huang C.R.L., Welch J., Gnanakkan V.P., Cornish T.C., Boeke J.D., Burns K.H. A map of DNA insertions in the NCI-60 human cancer cell panel. Mob. DNA 7:20-20(2016)
PubMed=28192450; DOI=10.1371/journal.pone.0171435 Fasterius E., Raso C., Kennedy S., Rauch N., Lundin P., Kolch W., Uhlen M., Al-Khalili Szigyarto C. A novel RNA sequencing data analysis method for cell line authentication. PLoS ONE 12:E0171435-E0171435(2017)
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=28683746; DOI=10.1186/s12943-017-0691-y Berg K.C.G., Eide P.W., Eilertsen I.A., Johannessen B., Bruun J., Danielsen S.A., Bjornslett M., Meza-Zepeda L.A., Eknaes M., Lind G.E., Myklebost O., Skotheim R.I., Sveen A., Lothe R.A. Multi-omics of 34 colorectal cancer cell lines - a resource for biomedical studies. Mol. Cancer 16:116-116(2017)
PubMed=28854368; DOI=10.1016/j.celrep.2017.08.010 Roumeliotis T.I., Williams S.P., Goncalves E., Alsinet C., Del Castillo Velasco-Herrera M., Aben N., Ghavidel F.Z., Michaut M., Schubert M., Price S., Wright J.C., Yu L., Yang M., Dienstmann R., Guinney J., Beltrao P., Brazma A., Pardo M., Stegle O., Adams D.J., Wessels L.F.A., Saez-Rodriguez J., McDermott U., Choudhary J.S. Genomic determinants of protein abundance variation in colorectal cancer cells. Cell Rep. 20:2201-2214(2017)
PubMed=29101300; DOI=10.15252/msb.20177701 Frejno M., Zenezini Chiozzi R., Wilhelm M., Koch H., Zheng R., Klaeger S., Ruprecht B., Meng C., Kramer K., Jarzab A., Heinzlmeir S., Johnstone E., Domingo E., Kerr D., Jesinghaus M., Slotta-Huspenina J., Weichert W., Knapp S., Feller S.M., Kuster B. Pharmacoproteomic characterisation of human colon and rectal cancer. Mol. Syst. Biol. 13:951-951(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、收到人結(jié)腸癌細胞HT-29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(lián)系。
2、收到人結(jié)腸癌細胞HT-29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(xiàn)細胞懸浮的情況,出現(xiàn)此狀態(tài)時,請不要打開細胞培養(yǎng)瓶,應(yīng)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(shù)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、收到人結(jié)腸癌細胞HT-29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、收到人結(jié)腸癌細胞HT-29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(zhuǎn)/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,人結(jié)腸癌細胞HT-29細胞形態(tài)已恢復(fù)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??梢苑?/span>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(nèi)離心,1000rpm/5min。棄上清,視細胞數(shù)量決定分瓶數(shù),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(jīng)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)。