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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(xùn)為一體的綜合化產(chǎn)業(yè)平臺,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)營原代細(xì)胞、細(xì)胞系、ELISA試劑盒、感受態(tài)細(xì)胞和HPLC檢測等科研產(chǎn)品與服務(wù)。我們秉承對用戶負(fù)責(zé)的態(tài)度,以對科研的高度嚴(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)!
更新時間:2021-05-24
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品牌 | 其他品牌 | 貨號 | BFN608006104 |
---|---|---|---|
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 | 人肺腺癌細(xì)胞Calu-3 | ||
貨物編碼 | BFN608006104 | ||
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 | 人肺腺癌細(xì)胞Calu-3細(xì)胞是從一名25歲的白人男性肺腺癌患者的胸水中分離建立的;該患者曾使用過環(huán)磷酰胺、博來霉素、阿霉素進行治療。該細(xì)胞接種至裸鼠可成瘤;可作轉(zhuǎn)染宿主。 該細(xì)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-55)。 | ||
注釋 | Part of: Cancer Cell Line Encyclopedia (CCLE) project. Part of: COSMIC cell lines project. Part of: KuDOS 95 cell line panel. Part of: MD Anderson Cell Lines Project. From: Memorial Sloan Kettering Cancer Center; New York; USA. Registration: Memorial Sloan Kettering Cancer Center Office of Technology Development; SK1980-533. Doubling time: 35 hours (in RPMI 1640 + 10% FBS), 45 hours (in ACL-3), 37 hours (in ACL-3+BSA) (PubMed=3940644). Microsatellite instability: Stable (MSS) (Sanger). Omics: Cell surface proteome. 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. Misspelling: CALV3; In Cosmic 1434969. | ||
STR信息 | Amelogenin:X;CSF1PO:11,12;D13S317:12;D16S539:12,14;D18S51:14,17;D19S433:12,13;D21S11:28,30;D2S1338:19;D3S1358:15,18;D5S818:11;D7S820:10,11;D8S1179:11,15;FGA:25;TH01:6,9.3;TPOX:8;vWA:16,17; | ||
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驗收細(xì)胞注意事項
1、收到人肺腺癌細(xì)胞Calu-3細(xì)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、收到人肺腺癌細(xì)胞Calu-3細(xì)胞,如包裝完好,請在顯微鏡下觀察細(xì)胞。,由于運輸過程中的問題,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,顯微鏡下觀察會出現(xiàn)細(xì)胞懸浮的情況,出現(xiàn)此狀態(tài)時,請不要打開細(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 小時左右,讓細(xì)胞先穩(wěn)定下,再于顯微鏡下觀察,此時多數(shù)細(xì)胞會重新貼附于瓶壁。如細(xì)胞仍不能貼壁,請用臺盼藍(lán)染色法鑒定細(xì)胞活力,如臺盼藍(lán)染色證實細(xì)胞活力正常請按懸浮細(xì)胞的方法處理。
3、收到人肺腺癌細(xì)胞Calu-3細(xì)胞后,請鏡下觀察細(xì)胞,用恰當(dāng)方式處理細(xì)胞。若懸浮的細(xì)胞較多,請離心收集細(xì)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(xì)胞,若細(xì)胞不多時 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、收到人肺腺癌細(xì)胞Calu-3細(xì)胞時如無異常情況 ,請在顯微鏡下觀察細(xì)胞密度,如為貼壁細(xì)胞,未超過80%匯合度時,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(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,以備不時之需。
6、24 小時后,人肺腺癌細(xì)胞Calu-3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,消化時間以具體細(xì)胞為準(zhǔn),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(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,有些生長較快的細(xì)胞則可以多傳幾瓶,以具體細(xì)胞和經(jīng)驗為準(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,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,請更換新鮮培養(yǎng)基培養(yǎng)。