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Tissue
肺部
Disease
腺癌
Metastatic site
淋巴结
Synonyms
H2009, H-2009, NCIH2009

Documents

NCI-H2009

Cell line · Product number 305283

US$550.00excl. tax and shipping
In stock
Lot
Recommended (freshest lot)
  • CoA
1

01General information

Description

NCI-H2009细胞系源自人类非小细胞肺癌(NSCLC),具体为腺癌类型。该细胞系在肺癌研究中被广泛应用,用于探究腺癌(NSCLC中最常见亚型)背后的分子与细胞机制。 NCI-H2009细胞在研究肺腺癌相关的基因突变、信号转导通路及治疗反应方面具有重要价值。

NCI-H2009细胞呈现上皮形态,表达肺腺癌特异性标志物,包括细胞角蛋白和癌胚抗原(CEA)。其携带NSCLC常见的遗传改变,如KRAS基因突变——该基因对细胞信号传导、生长及存活至关重要。 研究人员利用NCI-H2009细胞探索肺癌进展中的关键信号通路,如表皮生长因子受体(EGFR)、KRAS及磷脂酰肌醇-3激酶/Akt(PI3K/Akt)通路。该细胞系还应用于高通量药物筛选实验,以及化疗药物、靶向治疗和免疫疗法的临床前测试。此外,NCI-H2009细胞亦用于研究耐药机制及开发克服耐药性的策略。 NCI-H2009细胞系在肺腺癌研究中的重要性,彰显了其在深化肺癌生物学认知、为NSCLC患者开发新型高效治疗方案方面的关键作用。

Tissue
肺部
Disease
腺癌
Metastatic site
淋巴结
Synonyms
  • H2009
  • H-2009
  • NCIH2009

02Classification

Organism
Human
Sex
Female
Growth properties
Adherent
Biosafety level
1

03Characteristics

Morphology
上皮细胞

04Identifiers

Citation
NCI-H2009(赛通产品编号 305283)
NCBI TaxID
9606
Cellosaurus
CVCL_1514

05Expression and susceptibility

Mutational profile
突变:B2M,p.Met1Val (c.1A>G),杂合子;突变:B2M,p.Gln28Ter (c.82C>T),杂合子;突变:KRAS,p.Gly12Ala (c.35G>C),杂合子; 突变:TERT,c.1-124C>T (c.228C>T) (C228T);突变:TP53,p.Arg273Leu (c.818G>T),纯合子
Viruses
变异体:爱泼斯坦-巴氏病毒(EBV)

06Handling and culture

Culture medium

HITES培养基补充液

该细胞系的基础培养基为DF12。制备完全生长培养基时,需向基础培养基中添加以下组分:

  • 0.005 mg/ml 胰岛素
  • 0.01 mg/ml 转铁蛋白
  • 30 nM 亚硒酸钠(终浓度)
  • 10 nM 氢化可的松(终浓度)
  • 10 nM β-雌二醇(终浓度)
  • 额外添加2 mM L-谷氨酰胺(终浓度为4.5 mM)
  • 5% 胎牛血清(终浓度)
Incubation atmosphere

37°C, 5%CO2, 加湿环境。

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.

07Quality 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。

08Donor

Donor age
68岁
Ethnicity
欧洲