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WT-CLS1
Tissue
腎臟
Disease
肌細胞樣腫瘤
Synonyms
CLS1

Documents

WT-CLS1

Cell line · Product number 300379

The WT-CLS1 cell line was established from a primary Wilms’ tumor by CLS in 1998. However, the cells have rhabdoid characteristics, as demonstrated by E. Kun...

US$650.00excl. tax and shipping
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Lot
Recommended (freshest lot)
1

01General information

Description
WT-CLS1 細胞系由 CLS 於 1998 年從一例原發性威爾姆斯腫瘤中建立。然而,正如 E. Kunce Stroup 等人於 2017 年所示,該細胞具有橫紋樣特徵。 WT-CLS1 細胞對 miR-16 敏感,因此細胞周期蛋白 D 基因的表達會下降。此外,與真正的威爾姆斯腫瘤細胞不同,該細胞對 IGF1R 抑制展現出獨特的抗性。
Tissue
腎臟
Disease
肌細胞樣腫瘤
Synonyms
  • CLS1

02Classification

Organism
Human
Sex
Female
Growth properties
Adherent
Biosafety level
1

03Characteristics

Morphology
上皮樣
Cell type
B 淋巴母細胞
Tumourigenic
是的,在裸鼠身上。會形成由小細胞組成的腫瘤,其特徵與威爾姆斯腫瘤一致(異種移植模型可能無法完全反映威爾姆斯腫瘤的特徵,參見 E. Kunce Stroup 2017)。

04Identifiers

Citation
WT-CLS1(Cytion 型號 300379)
NCBI TaxID
9606
Cellosaurus
CVCL_5904

05Expression and susceptibility

Mutational profile
WT1 突變狀態:野生型;CTNNB1 突變狀態:野生型;無 LOH。
Viruses
HIV-1:陰性,HBV:陰性,HCV:陰性

06Handling and culture

Culture medium
IMDM,重量濃度:4.5 g/L 葡萄糖,重量濃度:4 mM L-谷氨醯胺,重量濃度:25 mM HEPES, w: 1.0 mM 丙酮酸鈉,w: 3.024 g/L NaHCO₃(Cytion 產品編號 820800a)
Incubation atmosphere

37°C,5% CO₂,加濕環境。

Supplements
在培養基中加入 10% 的胎牛血清(FBS)
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。
Seeding density
1 至 3 × 10⁵ 個細胞/cm²
Fluid renewal
每 3 至 4 天
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
5年
Ethnicity
高加索人