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Wilms10M
Tissue
腎臟
Disease
威爾姆斯腫瘤
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 細胞系是從一名威爾姆斯腫瘤(腎母細胞瘤)患者的轉移性肺結節中建立而來。 與其原發性腫瘤對應細胞系 Wilms10T 一樣,Wilms10M 細胞系的特徵在於 WT1 基因的純合性缺失,導致 WT1 蛋白完全缺失。WT1 對於腎臟的正常發育至關重要,其缺失與更具侵襲性的腫瘤行為有關,特別是在轉移性病變的情況下。 此外,Wilms10M 細胞在包含 IGF2 基因的 11p15 染色體區域呈現雜合性缺失(LOH),這進一步加劇了這些細胞的惡性特徵。

Wilms10M 細胞維持穩定的核型,除 WT1 區域的特定缺失外,並無重大染色體重排。此細胞系源自轉移性組織,對於研究驅動威爾姆斯腫瘤轉移的分子機制具有特別重要的價值。 這些細胞表現出間質特徵,表達如維曼汀(vimentin)等標記物,同時缺乏如細胞角蛋白(cytokeratin)等上皮標記物,這表明它們源自腫瘤的基質成分。

針對 Wilms10M 的研究主要聚焦於這些轉移性細胞中活躍的訊號傳導路徑。蛋白質體學分析顯示,數種受體酪胺酸激酶(RTKs)已活化,包括 IGF1R、PDGFRβ 及 AXL,這些受體參與促進細胞存活、增殖及轉移潛能。 下游的 MAPK 和 PI3K/AKT 訊號傳導路徑亦被活化,在維持 Wilms10M 細胞的侵襲性與轉移性表型方面扮演關鍵角色。鑒於其轉移性起源,Wilms10M 是理解威爾姆斯腫瘤轉移背後分子機制,以及開發針對轉移性疾病的標靶治療策略之重要模型。

Tissue
腎臟
Disease
威爾姆斯腫瘤
Applications
體外細胞培養模型。生化研究
Synonyms
  • Wilms10

02Classification

Organism
Human
Sex
Female
Growth properties
Adherent
Biosafety level
1

03Characteristics

Morphology
紡錘狀
Cell type
威爾姆斯細胞

04Identifiers

Citation
Wilms10M(Cytion 型號 300418)
NCBI TaxID
9606
Cellosaurus
CVCL_A5SL

05Expression and susceptibility

Mutational profile
WT1 突變狀態:在 del11p13 範圍內呈 WT1 純合性缺失。LOH:11p13 區域無 LOH,但 11p15 區域呈 UPD。CTNNB1 突變狀態:TCT 純合性缺失,p.DS45,3p 呈 UPD

06Handling and culture

Culture medium
MSCGM 套組(由 Lonza 提供)
Incubation atmosphere

37°C,5% CO₂,加濕環境。

Dissociation reagent
Accutase
Subculturing
從貼壁細胞中移除舊培養基,並用不含鈣和鎂的PBS洗滌細胞。T25培養瓶使用3-5毫升PBS,T75培養瓶則使用5-10毫升。 接著,用 Accutase 完全覆蓋細胞,T25 培養瓶使用 1–2 毫升,T75 培養瓶則使用 2.5 毫升。將細胞於室溫下孵育 8–10 分鐘,使其脫附。 孵育後,將細胞與 10 毫升培養基輕柔混合以使其重新懸浮,然後以 300xg 離心 3 分鐘。棄去上清液,將細胞重新懸浮於新鮮培養基中,並轉移至已預先加入新鮮培養基的新培養瓶中。
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

透過基於 PCR 的檢測及基於發光原理的支原體檢測方法,排除支原體污染的可能性。

為確保無細菌、真菌或酵母菌污染,每日均會對細胞培養物進行目視檢查。

Freeze medium
作為冷凍保存培養基,我們使用完全生長培養基(含 FBS)+ 10% DMSO 以確保解凍後的存活率,或使用 CM-1 (Cytion 型號 800100),該培養基含有經優化的滲透保護劑和代謝穩定劑,可提升細胞恢復率並減輕冷凍誘導的壓力。
Storage conditions

若需長期保存,請將試管置於溫度約為 −150 至 −196 °C 的氣相液氮中。僅在轉移至液氮之前,可將試管暫時存放於 −80 °C 環境中,作為短期過渡步驟。

Shipping conditions

冷凍保存的細胞系會以乾冰冷藏,並裝於經驗證的隔熱包裝中,包裝內含有足夠的冷媒,以確保在運輸過程中溫度維持在約 −78 °C。收到貨物後,請立即檢查容器,並盡快將試管轉移至適當的儲存場所。

08Donor

Donor age
2年
Ethnicity
高加索人