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NCI-H196
Tissue
肺部
Disease
肺小细胞癌
Metastatic site
胸腔积液
Synonyms
NCI-H196, H-196, NCIH196

Documents

NCI-H196

Cell line · Product number 300390

NCI-H196 is a small-cell lung cancer (SCLC) cell line used to study the mechanisms of cancer progression, chemotherapy resistance, and cellular responses ...

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01General information

Description

NCI-H196 是一种小细胞肺癌(SCLC)细胞系,用于研究癌症进展机制、化疗耐药性和细胞对氧化应激的反应。对 NCI-H196 的研究表明,它对吡咯烷二硫代氨基甲酸盐(PDTC)的细胞毒性作用非常敏感,而 PDTC 是一种促氧化剂。PDTC 可诱导 S 期细胞周期停滞,并以剂量依赖的方式显著降低 NCI-H196 细胞的存活率。这种细胞毒性归因于氧化应激的诱导,表现为活性氧(ROS)的增加和氧化应激相关基因表达的变化。添加 N-乙酰-L-半胱氨酸(NAC)等抗氧化剂可有效逆转 PDTC 诱导的细胞毒性,从而证实氧化应激在细胞死亡中的作用。

进一步的研究表明,PDTC 可增强顺铂的细胞毒性,顺铂是治疗 SCLC 的一线化疗药物。将低剂量的顺铂与无毒浓度的 PDTC 结合使用,可在 NCI-H196 细胞中产生协同细胞毒性。据信,这种联合疗法之所以有效,是因为 PDTC 下调了与顺铂耐药性有关的铜外排转运体 ATP7A。通过抑制 ATP7A,PDTC 可增加细胞内的铜,并使 NCI-H196 细胞对顺铂敏感,从而突出了其作为 SCLC 辅助疗法的潜力。

Tissue
肺部
Disease
肺小细胞癌
Metastatic site
胸腔积液
Applications
三维细胞培养、癌症研究
Synonyms
  • NCI-H196
  • H-196
  • NCIH196

02Classification

Organism
Human
Sex
Male
Growth properties
Adherent
Biosafety level
1

03Identifiers

Citation
NCI-H196(Cytion 目录号 300390)
NCBI TaxID
9606
Cellosaurus
CVCL_1509

04Handling 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。
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.

05Quality 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。

06Donor

Donor age
68岁
Ethnicity
欧洲