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Wilms10M
Tissue
肾脏
Disease
Wilms 肿瘤
Synonyms
Wilms10

Documents

Wilms10M

Cell line · Product number 300418

The Wilms10M cell line was established from a metastatic lung nodule of a patient with Wilms tumor (nephroblastoma). Like its primary tumor counterpart, W...

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01General information

Description

Wilms10M 细胞系是从一名 Wilms 肿瘤(肾母细胞瘤)患者的转移性肺结节中建立的。与原发肿瘤 Wilms10T 细胞系一样,Wilms10M 细胞系的特点是 WT1 基因同源缺失,导致 WT1 蛋白完全缺失。WT1 是肾脏正常发育所必需的,其缺失与更具侵袭性的肿瘤行为有关,尤其是在转移情况下。此外,Wilms10M 细胞在包括 IGF2 基因在内的 11p15 染色体区域表现出杂合性缺失(LOH),进一步加剧了这些细胞的恶性特性。

Wilms10M 细胞保持稳定的核型,除了 WT1 区域的特异性缺失外,没有重大的染色体重排。这种细胞系来源于转移组织,对研究驱动 Wilms 肿瘤转移的分子机制特别有价值。这些细胞表现出间充质特征,表达波形蛋白等标志物,但缺乏细胞角蛋白等上皮标志物,这表明它们来自肿瘤的基质成分。

对 Wilms10M 的研究主要集中在这些转移细胞中活跃的信号通路。蛋白质组分析表明,包括 IGF1R、PDGFRβ 和 AXL 在内的多种受体酪氨酸激酶(RTK)被激活,它们参与促进细胞存活、增殖和转移潜能。下游的 MAPK 和 PI3K/AKT 信号通路也被激活,在维持 Wilms10M 细胞的侵袭和转移表型方面发挥着关键作用。鉴于其转移起源,Wilms10M 是了解 Wilms 肿瘤转移的分子事件和开发针对转移性疾病的靶向治疗策略的重要模型。

Tissue
肾脏
Disease
Wilms 肿瘤
Applications
体外细胞培养模型。生化研究
Synonyms
  • Wilms10

02Classification

Organism
Human
Sex
Female
Growth properties
Adherent
Biosafety level
1

03Characteristics

Morphology
纺锤形
Cell type
Wilms 细胞

04Identifiers

Citation
Wilms10M(Cytion 目录号 300418)
NCBI TaxID
9606
Cellosaurus
CVCL_A5SL

05Expression and susceptibility

Mutational profile
WT1 基因突变情况:在 del11p13 中为同卵 del WT1。LOH:11p13 无,但 11p15 有 UPD。 CTNNB1 突变状态:同基因 del TCT,p.DS45,UPD 3p

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。
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