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Wilms1
Tissue
肾脏
Synonyms
Wilms1-2l

Documents

Wilms1

Cell line · Product number 300411

The Wilms1 cell line was derived from a primary Wilms tumor sample obtained from a patient presenting with large bilateral kidney tumors, indicative of Wi...

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01General information

Description

Wilms1 细胞系来源于一个原发性 Wilms 肿瘤样本,该样本取自一名患有双侧肾脏大肿瘤的患者,表明该患者患有 Wilms 肿瘤(一种小儿肾母细胞瘤)。该细胞系的 WT1 基因存在同卵无义突变(c.149 C>A,p.S50X),导致 WT1 蛋白截短且无功能。WT1 基因对肾脏的发育和功能至关重要,在 Wilms 肿瘤中经常发生突变,尤其是在那些表现出异位间质分化的基质亚型中。因此,Wilms1 细胞是研究肿瘤生物学中 WT1 功能缺失后果的独特体外模型。

Wilms1 细胞系保持稳定的核型,没有明显的染色体异常,可以进行可靠的长期培养。这些细胞表现出间质表型,其特点是表达波形蛋白,而不表达细胞角蛋白等上皮标记物,这与其基质来源一致。此外,该细胞系还表现出有限但显著的间充质分化能力,包括在适当条件下分化成肌肉样细胞的能力。这使得 Wilms1 成为研究间质分化分子机制及其在 Wilms 肿瘤发病机制中失调的宝贵工具。

Wilms1 还被用于研究参与肿瘤进展的关键信号通路的激活状态。蛋白质组分析表明,Wilms1 细胞表现出多种受体酪氨酸激酶的磷酸化和激活,包括表皮生长因子受体和表皮生长因子受体β,以及下游的 MAPK 信号通路。这些发现凸显了 Wilms1 细胞系在探索 Wilms 肿瘤靶向治疗方法中的重要性,因为它能剖析这些通路在癌细胞存活、增殖和分化中的作用。

Tissue
肾脏
Applications
体外细胞培养模型。生化研究
Synonyms
  • Wilms1-2l

02Classification

Organism
Human
Sex
Female
Growth properties
Adherent
Biosafety level
1

03Characteristics

Morphology
纺锤形
Cell type
Wilms 细胞
Tumourigenic
是,在裸鼠体内。形成与 Wilms 肿瘤一致的带有小细胞的肿瘤(异种移植物可能并不完全代表 Wilm 肿瘤,参见 E. Kunce Stroup,2017 年)
Karyotype
46、正常
Doubling time
24 小时

04Identifiers

Citation
Wilms1(Cytion 目录编号 300411)
NCBI TaxID
9606
Cellosaurus
CVCL_A5SC

05Expression and susceptibility

Receptors expressed
受体酪氨酸激酶 表皮生长因子受体、EphA7、表皮生长因子受体α、表皮生长因子受体1、表皮生长因子受体β、AxL
Mutational profile
WT1 突变状态:同基因 c. 149 C>A, p.S50x, LOH: 11p11-11pter, CTNNB1 突变状态:异基因 TCT>TTT, p.S45F
Viruses
HIV-1: 阴性、HBV: 阴性、HCV: 阴性

06Handling and culture

Culture medium
MSCGM 试剂盒(龙沙公司提供)
Incubation atmosphere

37°C, 5%CO2, 加湿环境。

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。
Seeding density
1 × 10⁴ 个细胞/平方厘米
Fluid renewal
每周 1 至 2 次
Post-thaw recovery
快速
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
2 年
Ethnicity
高加索人