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NCI-H1048
Tissue
肺部
Disease
小细胞癌
Metastatic site
胸腔积液
Synonyms
H1048, H-1048, NCIH1048

Documents

NCI-H1048

Cell line · Product number 305595

NCI-H1048 is a human small cell lung carcinoma (SCLC) cell line derived from a lung tumor in an adult patient and is widely used as a model of neuroendocr...

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01General information

Description

NCI-H1048 是一种源自成年患者肺部肿瘤的人小细胞肺癌(SCLC)细胞系,被广泛用作神经内分泌肺癌的模型。 小细胞肺癌以生长迅速、早期转移和与神经内分泌分化密切相关为特征,而 NCI-H1048 反映了其中许多特征。这些细胞通常以悬浮或松散粘附的簇状形式生长,并表现出与 SCLC 一致的形态,包括细胞体积小、呈圆形且核质比高。

在分子水平上,NCI-H1048 表现出小细胞肺癌的特征性表现,包括关键抑癌基因通路(如 TP53 和 RB1)的改变,这些基因在该疾病中通常处于失活状态。 该细胞系表达神经内分泌标志物,包括与激素分泌和神经元分化相关的蛋白质,使其成为研究神经内分泌信号传导和肿瘤生物学的重要模型。与其他小细胞肺癌模型类似,它也可能表现出参与增殖和生存的致癌驱动基因的扩增或过表达,从而促使其呈现侵袭性表型。

NCI-H1048 用于针对小细胞肺癌发病机制、药物敏感性及耐药机制的研究。在以初始治疗反应良好但随后迅速复发为特征的疾病背景下,该细胞系对于评估化疗药物和靶向治疗具有特别重要的价值。 该细胞系还被用于肿瘤细胞可塑性、神经内分泌分化以及高通量药物筛选的研究。然而,与许多小细胞肺癌模型一样,不同数据集间的详细突变特征可能存在差异,因此对于需要精确基因组信息的实验,建议进行额外的分子特征分析。

Tissue
肺部
Disease
小细胞癌
Metastatic site
胸腔积液
Synonyms
  • H1048
  • H-1048
  • NCIH1048

02Classification

Organism
Human
Sex
Female
Growth properties
Adherent
Biosafety level
1

03Characteristics

Morphology
上皮样
MSI status
不稳定(MSI 高)

04Identifiers

Citation
NCI-H1048(Cytion 目录号 305595)
NCBI TaxID
9606
Cellosaurus
CVCL_1453

05Handling and culture

Culture medium
DMEM:Ham's F12(1:1),w: 3.1 g/L 葡萄糖,w: 2.5 mM L-谷氨酰胺,w: 15 mM HEPES,w: 0.5 mM 丙酮酸钠,w: 1.2 g/L NaHCO3(Cytion 文章编号 820400a)。
Incubation atmosphere

37°C, 5%CO2, 加湿环境。

Supplements
在培养基中添加 5% FBS、0.005 mg/mL 胰岛素、0.01 mg/mL 转铁蛋白、30nM 亚硒酸钠、10 nM 氢化可的松、10 nM beta-estradiol
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。
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. Medium

    HITES medium supplemented with 5% fetal bovine serum: The base medium for this cell line is DMEM:F12 Medium (Catalog No. 820400a). To make the complete growth medium, add the following components to the base medium:

    • 0.005 mg/ml Insulin
    • 0.01 mg/ml Transferrin
    • 30 nM Sodium selenite (final conc.)
    • 10 nM Hydrocortisone (final conc.)
    • 10 nM beta-estradiol (final conc.)
    • extra 2 mM L-glutamine (for final conc. of 4.5 mM)
    • 5% fetal bovine serum (final conc.)
  7. Centrifuge the mixture at 300 x g for 3 minutes to separate the cells and carefully discard the supernatant containing residual freezing medium.

  8. 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.

  9. 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

Donor age
53 年
Ethnicity
非洲裔美国人