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CaSki 細胞
Tissue
子宮頸
Disease
癌
Metastatic site
子宮頸
Synonyms
Ca-Ski, Ca Ski, Caski, CASKI

Documents

CaSki 細胞

Cell line · Product number 300145

CaSki is a cell line exhibiting epithelial morphology, isolated from the cervix of a 40-year-old White female patient with epidermoid carcinoma. The estab...

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01General information

Description

CaSki 是一種呈現上皮形態的細胞系,取自一名 40 歲白人女性患者(患有鱗狀細胞癌)的子宮頸。 該細胞系的建立為子宮頸癌研究提供了關鍵模型,特別是在 HPV 介導的致癌機制方面。CaSki 細胞的特徵在於其能夠複製 HPV16 DNA,該 DNA 會整合至宿主基因組中,有助於深入了解病毒的生命週期及其在惡性轉化中的作用。

這些細胞是癌症研究中不可或缺的資源,特別是針對HPV相關子宮頸癌病理機制的研究。 CaSki 細胞中存在高風險型 HPV16,有助於探索病毒致癌基因的功能,尤其是 E6 和 E7 蛋白,以及它們與細胞腫瘤抑制路徑(包括涉及 p53 和 pRB 的路徑)之間的相互作用。這一點使 CaSki 細胞在評估潛在治療靶點以及開發針對 HPV 誘導性惡性腫瘤的干預措施方面具有無可替代的價值。

Tissue
子宮頸
Disease
癌
Metastatic site
子宮頸
Synonyms
  • Ca-Ski
  • Ca Ski
  • Caski
  • CASKI

02Classification

Organism
Human
Sex
Female
Growth properties
Adherent
Biosafety level
2

03Characteristics

Morphology
上皮樣
Cell type
鱗狀細胞癌

04Identifiers

Citation
CaSki(Cytion 型號 300145)
NCBI TaxID
9606
Cellosaurus
CVCL_1100

05Expression and susceptibility

Isoenzymes
G6PD, B
Products
人絨毛膜促性腺激素(hCG)的β亞單位,腫瘤相關抗原

06Handling and culture

Culture medium
RPMI 1640,含 2.0 mM 穩定谷氨醯胺,含 2.0 g/L NaHCO₃(Cytion 產品編號 820700a)
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 × 10⁴ 個細胞/cm² 將在 3 至 4 天內形成密合的單層細胞。
Fluid renewal
每週 2 至 3 次
Post-thaw recovery
解凍後,將細胞以 5 × 10⁴ 個細胞/cm² 的密度接種於培養皿中,並讓細胞從冷凍過程中恢復,並附著至少 48 小時。
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
40年
Ethnicity
高加索人