人非小細胞肺癌細胞NCI-H358
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| 供貨周期 | 現(xiàn)貨 | 規(guī)格 | T25培養(yǎng)瓶x1 1.5ml凍存管x2 |
|---|---|---|---|
| 貨號 | BFN60800666 | 應用領(lǐng)域 | 醫(yī)療衛(wèi)生,生物產(chǎn)業(yè) |
| 主要用途 | 僅供科研 |
細胞名稱 | 人非小細胞肺癌細胞NCI-H358 |
| |
貨物編碼 | BFN60800666 | ||
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 | 人非小細胞肺癌細胞NCI-H358細胞于1981年從一位開始化療之前的患者的腫瘤組織中分離建株。超微結(jié)構(gòu)研究表明細胞質(zhì)中有Clara細胞的特征結(jié)構(gòu)細胞表達主要的肺表面結(jié)合蛋白SP-A的蛋白和RNA。不表達SP-B和SP-C。他們在軟瓊脂中的克隆形成效率為0.83%。 | ||
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: NCI RAS program mutant KRAS cell line panel. Doubling time: 38 hours (in RPMI 1640 + 10% FBS), 76 hours (in ACL-3), 60 hours (in ACL-3 + BSA) (PubMed=3940644); 38 hours (ECACC). Microsatellite instability: Stable (MSS) (Sanger). Omics: Deep exome analysis. Omics: Deep phosphoproteome analysis. Omics: Deep proteome analysis. Omics: Deep RNAseq analysis. Omics: DNA methylation analysis. Omics: Protein expression by reverse-phase protein arrays. Omics: SNP array analysis. Omics: Transcriptome analysis. Caution: We are not certain that NCI-H358 and NCI-H358M are identical. Misspelling: H1358; In PubMed=1311061 table 1. | ||
STR信息 | Amelogenin:X,Y；CSF1PO:11，12；D13S317:8，12；D16S539:12，13；D18S51:14；D19S433:13，14；D21S11:28，30；D2S1338:17，23；D3S1358:14，18；D5S818:10,12；D7S820:10，11；D8S1179:13，14；FGA:20,21；TH01:6；TPOX:8，9；vWA:17 | ||
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驗收細胞注意事項
1、收到人非小細胞肺癌細胞NCI-H358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、收到人非小細胞肺癌細胞NCI-H358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、收到人非小細胞肺癌細胞NCI-H358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、收到人非小細胞肺癌細胞NCI-H358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，人非小細胞肺癌細胞NCI-H358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?？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)。
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