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NCI-H1373
Tissue
肺部
Disease
腺癌
Synonyms
H1373, H-1373, NCIH1373

Documents

NCI-H1373

Cell line · Product number 305597

The NCI-H1373 cell line is a human non-small cell lung carcinoma (NSCLC) model harboring the KRAS G12C mutation, which is frequently associated with oncog...

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01General information

Description

NCI-H1373细胞系是一种携带KRAS G12C突变的人类非小细胞肺癌(NSCLC)模型,该突变常与肺腺癌的致瘤过程相关。 该突变会导致下游信号传导通路(如丝裂原活化蛋白激酶(MAPK)和磷脂酰肌醇3激酶(PI3K)通路)的持续激活,从而促进肿瘤的增殖和存活。 NCI-H1373 被广泛用于研究,以评估针对 KRAS G12C 及相关致癌通路的新型治疗药物的疗效。

在临床前研究中,NCI-H1373已被用于评估新兴的KRAS G12C抑制剂(如ASP2453)的活性。该抑制剂表现出对突变型KRAS蛋白的强效、选择性且共价结合能力,可在体外抑制KRAS介导的信号传导和肿瘤细胞增殖。 该细胞系还被用于异种移植模型,在该模型中,ASP2453诱导了显著的肿瘤退缩。这一活性凸显了 NCI-H1373 在理解新型 KRAS 抑制剂的治疗潜力和作用机制方面的实用价值。

此外,NCI-H1373已被用于全基因组CRISPR敲除筛选,以探索PARP7抑制剂耐药的机制。 研究结果表明,芳香烃受体(AHR)信号传导的缺失与PARP7抑制剂耐药性相关,这为潜在的联合治疗策略提供了依据。此类研究证明了该细胞系在识别耐药通路以及优化针对KRAS G12C突变型非小细胞肺癌(NSCLC)的靶向治疗方案方面的作用。

Tissue
肺部
Disease
腺癌
Synonyms
  • H1373
  • H-1373
  • NCIH1373

02Classification

Organism
Human
Sex
Male
Growth properties
Adherent
Biosafety level
1

03Characteristics

Morphology
上皮样

04Identifiers

Citation
NCI-H1373(Cytion 产品编号 305597)
NCBI TaxID
9606
Cellosaurus
CVCL_1465

05Expression and susceptibility

Mutational profile
突变:KRAS,p.Gly12Cys (c.34G>T),纯合子;突变:TP53,p.Pro47Leu (c.140C>T),纯合子

06Handling 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 TrypLE Express 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 毫升培养基轻轻混匀细胞使其重悬,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
56 岁
Ethnicity
非洲裔美国人