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NCI-H209 细胞
Tissue
肺部
Disease
小细胞癌
Metastatic site
骨髓
Synonyms
H209, H-209, NCIH209

Documents

NCI-H209 细胞

Cell line · Product number 300183

The NCI-H209 cell line was derived by A.F. Gazdar and associates in 1979 from the bone marrow of a patient with small cell cancer of the lung. The bone marro...

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1

01General information

Description
NCI-H209 细胞系是 A.F. Gazdar 及其同事于 1979 年从一名肺小细胞癌患者的骨髓中提取的。骨髓标本是在治疗前采集的。该细胞系是一种典型的 SCLC 细胞系,能表达四种生化标志物(神经元特异性烯醇化酶、脑肌酸激酶同工酶、L-DOPA 脱羧酶和蚕豆蛋白样免疫反应)的高水平。C-myc DNA 序列未被扩增。未发现严重的 DNA 结构异常。这是一种在悬浮液中生长为大型聚集体的细胞系。只有聚集体才有活力,但无法测量有意义的活力百分比。培养基中通常含有大量细胞碎片。该细胞表达一种不磷酸化的 RB1 异常形式,这显然是由于密码子 706 处的单点突变(Cys-> Phe)造成的。
Tissue
肺部
Disease
小细胞癌
Metastatic site
骨髓
Synonyms
  • H209
  • H-209
  • NCIH209

02Classification

Organism
Human
Sex
Male
Growth properties
Suspension
Biosafety level
1

03Characteristics

Morphology
上皮样
Tumourigenic
是,在裸鼠体内形成具有典型 SCLC 组织学的可移植肿瘤

04Identifiers

Citation
NCI-H209(Cytion 目录号 300183)
NCBI TaxID
9606
Cellosaurus
CVCL_1525

05Expression and susceptibility

Protein expression
P53 阴性
Isoenzymes
G6PD,B,PGM1,1-2,PGM3,1,ES-D,1,Me-2,0,AK-1,1,GLO-1,1-2,表型频率乘积 = 0.0624
Products
与正常肺部相比,该品系能产生正常数量的 p53 mRNA。

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
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。
Seeding density
1 × 10⁵ 个细胞/毫升
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
55 岁
Ethnicity
高加索人