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NCI-H446
Tissue
肺部
Disease
肺小细胞癌
Metastatic site
胸腔积液
Synonyms
NCI-H446, H-446, NCI-446, NCIH446

Documents

NCI-H446

Cell line · Product number 305049

This cell line was established in 1982 by D. Carney, A.F. Gazdar and associates from the pleural fluid of a patient with small cell cancer of the lung. The o...

US$395.00excl. tax and shipping
In stock
Lot
Recommended (freshest lot)
  • CoA
1

01General information

Description
该细胞系由 D. Carney、A.F. Gazdar 及其同事于 1982 年从一名肺小细胞癌患者的胸腔积液中建立。最初的肿瘤形态并不具有小细胞肺癌的特征。该细胞系在生物化学和形态上属于变异型小细胞肺癌,能表达神经元特异性烯醇化酶和脑肌酸激酶同工酶。在该细胞系中未检测到 L-DOPA脱羧酶、加压素、血管加压素、催产素或胃泌素释放肽。该细胞系的 c-myc DNA 扩增程度高出 20 倍,c-myc RNA 扩增程度高出 15 倍。该细胞系最初是在无血清的 RPMI 1640 培养基中繁殖的,培养基中补充了 10 nM 的氢化可的松、5 微克/毫升的胰岛素、10 微克/毫升的转铁蛋白、10 nM 的 17-beta-estradiol 和 30 nM 的亚硒酸钠。细胞可形成非典型小细胞肺癌组织学的可移植肿瘤。
Tissue
肺部
Disease
肺小细胞癌
Metastatic site
胸腔积液
Synonyms
  • NCI-H446
  • H-446
  • NCI-446
  • NCIH446

02Classification

Organism
Human
Sex
Male
Growth properties
Adherent
Biosafety level
1

03Characteristics

Morphology
上皮样
Tumourigenic
是的,在裸鼠体内(细胞形成可移植的非典型 SCLC 组织学肿瘤)。

04Identifiers

Citation
NCI-H446(Cytion 目录号 305049)
NCBI TaxID
9606
Cellosaurus
CVCL_1562

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、2.5 克/升葡萄糖、10 毫摩尔 HEPES 和 1.0 毫摩尔丙酮酸钠。
Dissociation reagent
Accutase
Subculturing
将悬浮细胞收集到 15 毫升试管中,用不含钙和镁的 PBS(T25 烧瓶用 3-5 毫升,T75 烧瓶用 5-10 毫升)轻轻清洗粘附的细胞。涂抹 Accutase(T25 烧瓶 1-2 毫升,T75 烧瓶 2.5 毫升),确保完全覆盖细胞层。让细胞在室温下孵育 10 分钟。孵育后,将悬浮细胞和粘附细胞混合并离心。离心后,小心重悬细胞团,并将细胞悬浮液转移到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

Donor age
61 年
Ethnicity
欧洲