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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(hào)	BFN60800666	應(yīng)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(xì)胞名稱(chēng)	人非小細(xì)胞肺癌細(xì)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(xì)胞數(shù)量	1x10^6	1x10^6
保存溫度	37℃	-198℃
運(yùn)輸方式	常溫保溫運(yùn)輸	干冰運(yùn)輸
安全等級(jí)	1
用途限制	僅供科研用途 1類(lèi)

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(jiǎn)介	人非小細(xì)胞肺癌細(xì)胞NCI-H358細(xì)胞于1981年從一位開(kāi)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(xì)胞質(zhì)中有Clara細(xì)胞的特征結(jié)構(gòu)細(xì)胞表達(dá)主要的肺表面結(jié)合蛋白SP-A的蛋白和RNA。不表達(dá)SP-B和SP-C。他們?cè)谲洯傊械目寺⌒纬尚?/strong>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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Peripheral airway cell differentiation in human lung cancer cell lines. Cancer Res. 50:5481-5487(1990) PubMed=1311061 Mitsudomi T., Steinberg S.M., Nau M.M., Carbone D., D'Amico D., Bodner S., Oie H.K., Linnoila R.I., Mulshine J.L., Minna J.D., Gazdar A.F. p53 gene mutations in non-small-cell lung cancer cell lines and their correlation with the presence of ras mutations and clinical features. Oncogene 7:171-180(1992) PubMed=1565469 Bodner S.M., Minna J.D., Jensen S.M., D'Amico D., Carbone D.P., Mitsudomi T., Fedorko J., Buchhagen D.L., Nau M.M., Gazdar A.F., Linnoila R.I. Expression of mutant p53 proteins in lung cancer correlates with the class of p53 gene mutation. Oncogene 7:743-749(1992) PubMed=8806092; DOI=10.1002/jcb.240630505 Phelps R.M., Johnson B.E., Ihde D.C., Gazdar A.F., Carbone D.P., McClintock P.R., Linnoila R.I., Matthews M.J., Bunn P.A. Jr., Carney D.N., Minna J.D., Mulshine J.L. NCI-Navy Medical Oncology Branch cell line data base. J. Cell. 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驗(yàn)收細(xì)胞注意事項(xiàng)

1、收到人非小細(xì)胞肺癌細(xì)胞NCI-H358細(xì)胞，請(qǐng)查看瓶子是否有破裂，培養(yǎng)基是否漏出，是否渾濁，如有請(qǐng)盡快聯(lián)系。

2、收到人非小細(xì)胞肺癌細(xì)胞NCI-H358細(xì)胞，如包裝完好，請(qǐng)?jiān)陲@微鏡下觀(guān)察細(xì)胞。,由于運(yùn)輸過(guò)程中的問(wèn)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(lái)，顯微鏡下觀(guān)察會(huì)出現(xiàn)細(xì)胞懸浮的情況，出現(xiàn)此狀態(tài)時(shí)，請(qǐng)不要打開(kāi)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(shí)左右，讓細(xì)胞先穩(wěn)定下，再于顯微鏡下觀(guān)察，此時(shí)多數(shù)細(xì)胞會(huì)重新貼附于瓶壁。如細(xì)胞仍不能貼壁，請(qǐng)用臺(tái)盼藍(lán)染色法鑒定細(xì)胞活力，如臺(tái)盼藍(lán)染色證實(shí)細(xì)胞活力正常請(qǐng)按懸浮細(xì)胞的方法處理。

3、收到人非小細(xì)胞肺癌細(xì)胞NCI-H358細(xì)胞后，請(qǐng)鏡下觀(guān)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(qǐng)離心收集細(xì)胞，接種到一個(gè)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(jìn)口胎牛血清。剛接到細(xì)胞，若細(xì)胞不多時(shí) 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、收到人非小細(xì)胞肺癌細(xì)胞NCI-H358細(xì)胞時(shí)如無(wú)異常情況，請(qǐng)?jiān)陲@微鏡下觀(guān)察細(xì)胞密度，如為貼壁細(xì)胞，未超過(guò)80%匯合度時(shí)，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(guò) 80%匯合度時(shí)，請(qǐng)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(guò)的瓶子里，存放于 4℃冰箱，以備不時(shí)之需。

6、24 小時(shí)后，人非小細(xì)胞肺癌細(xì)胞NCI-H358細(xì)胞形態(tài)已恢復(fù)并貼滿(mǎn)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(zhǎng)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(shí)間以具體細(xì)胞為準(zhǔn)，一般 1-3 分鐘，不超過(guò) 5 分鐘?？梢苑?/span>入37℃培養(yǎng)箱消化。輕輕晃動(dòng)瓶壁，見(jiàn)細(xì)胞脫落下來(lái)，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(guò)稀，有些生長(zhǎng)較快的細(xì)胞則可以多傳幾瓶，以具體細(xì)胞和經(jīng)驗(yàn)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。