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NCI-H460
Tissue
肺部
Disease
肺大细胞癌
Metastatic site
胸腔积液
Synonyms
NCI-H460, NCI.H460, H-460, NCIH460, NCI-HUT-460, NCI-460

Documents

NCI-H460

Cell line · Product number 305020

NCI-H460, also known as H460, was derived from a male patient with large cell lung carcinoma. NCI-H460 cells are adherent cells growing twice as fast than th...

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Lot
Recommended (freshest lot)
  • CoA
1

01General information

Description
NCI-H460,又称 H460,来源于一名男性大细胞肺癌患者。NCI-H460 细胞是一种粘附细胞,其生长速度是 A549 细胞的两倍,在添加了 10% FBS 的 RPMI 1640 细胞中翻倍时间为 33 小时。它们可以在体外和体内模型(包括裸鼠)中形成肿瘤。研究表明,NCI-H460 细胞表达 p53 mRNA 的水平很高,可与正常肺组织媲美,但没有出现严重的 DNA 结构异常。它们的角蛋白和波形蛋白染色呈阳性,但神经丝三联蛋白染色呈阴性。同工酶分析表明,HPRT 定位于这些非小细胞肺癌细胞系的表面。AK-1、ES-D 和 Me-2 同工酶在 1 级表达,而 G6PD 和 PGM1 及 PGM3 同工酶分别在 B 级和 1-2 级表达。这些细胞的核型为低倍体,染色体数目模数为 57,从 53 到 65 不等。它们的 p53 mRNA 高表达水平使其成为研究非小细胞肺癌分子机制的合适模型。
Tissue
肺部
Disease
肺大细胞癌
Metastatic site
胸腔积液
Synonyms
  • NCI-H460
  • NCI.H460
  • H-460
  • NCIH460
  • NCI-HUT-460
  • NCI-460

02Classification

Organism
Human
Sex
Male
Growth properties
Adherent
Biosafety level
1

03Characteristics

Morphology
上皮细胞
Tumourigenic
是

04Identifiers

Citation
H-460(Cytion 目录号 305020)
NCBI TaxID
9606
Cellosaurus
CVCL_0459

05Handling and culture

Culture medium
RPMI 1640,w:2.0 mM 稳定谷氨酰胺,w:2.0 g/L NaHCO3(Cytion 文章编号 820700a)。
Incubation atmosphere

37°C, 5%CO2, 加湿环境。

Supplements
在培养基中添加 10% FBS
Dissociation reagent
Accutase
Subculturing
去除粘附细胞上的旧培养基,用不含钙和镁的 PBS 冲洗。T25 烧瓶用 3-5 毫升 PBS,T75 烧瓶用 5-10 毫升。然后用 Accutase 完全覆盖细胞,T25 烧瓶用 1-2 毫升,T75 烧瓶用 2.5 毫升。让细胞在室温下孵育 8-10 分钟,使其分离。孵育后,用 10 毫升培养基轻轻混合细胞使其重悬,然后用 300xg 离心 3 分钟。弃去上清液,用新鲜培养基重悬细胞,并将其转移到已装有新鲜培养基的新烧瓶中。
Fluid renewal
每周 2 至 3 次
Thawing and culturing
  1. Confirm that the vial remains deeply frozen upon delivery, as cells are shipped on dry ice to maintain optimal temperatures during transit.

  2. Upon receipt, either store the cryovial immediately at temperatures below -150°C to ensure the preservation of cellular integrity, or proceed to step 3 if immediate culturing is required.

  3. For immediate culturing, swiftly thaw the vial by immersing it in a 37°C water bath with clean water and an antimicrobial agent, agitating gently for 40-60 seconds until a small ice clump remains.

  4. Perform all subsequent steps under sterile conditions in a flow hood, disinfecting the cryovial with 70% ethanol before opening.

  5. Carefully open the disinfected vial and transfer the cell suspension into a 15 ml centrifuge tube containing 8 ml of room-temperature culture medium, mixing gently.

  6. Centrifuge the mixture at 300 x g for 3 minutes to separate the cells and carefully discard the supernatant containing residual freezing medium.

  7. Gently resuspend the cell pellet in 10 ml of fresh culture medium. For adherent cells, divide the suspension between two T25 culture flasks; for suspension cultures, transfer all the medium into one T25 flask to promote effective cell interaction and growth.

  8. Adhere to established subculture protocols for continued growth and maintenance of the cell line, ensuring reliable experimental outcomes.

06Quality control and cryopreservation

Sterility

使用基于 PCR 的检测方法和基于发光的支原体检测方法排除支原体污染。

为确保没有细菌、真菌或酵母菌污染,每天都要对细胞培养物进行目视检查。

Freeze medium
我们使用完全生长培养基(包括 FBS)+10% DMSO 或 CM-1(Cytion 商品目录编号 800100)作为冷冻保存培养基,以获得足够的解冻后存活率,CM-1(Cytion 商品目录编号 800100)含有优化的渗透保护剂和代谢稳定剂,可提高恢复能力并减少冷冻引起的应激。
Storage conditions

如需长期保存,可将样品瓶放入气相液氮中,温度约为 -150 至 -196 ℃。在-80 °C下保存只能作为转入液氮前的短暂过渡。

Shipping conditions

冷冻保存的细胞系用干冰装运,并用经过验证的隔热包装和足够的制冷剂在整个运输过程中保持约 -78 °C 的温度。收到货后,应立即检查容器,并立即将小瓶转移到适当的储藏室。

07Donor

Ethnicity
欧洲