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U14
Disease
子宮頸癌

Documents

U14

Cell line · Product number 305509

The U14 cell line is a murine cervical squamous cell carcinoma model that was originally established by treating the uterine cervix of mice with 20-methyl...

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01General information

Description

U14 細胞系是一種小鼠子宮頸鱗狀細胞癌模型,最初是透過以 20-甲基膽蒽處理小鼠子宮頸而建立的。U14 細胞已廣泛應用於癌症研究,特別是用於研究腫瘤侵襲、轉移及藥物篩選。 當將其植入小鼠體內時,這些細胞展現出極高的轉移潛能,其淋巴轉移率與肺轉移率分別達 95% 及 80%。在體外實驗中,這些細胞呈現上皮細胞特徵,包括細胞角蛋白染色呈陽性,且具有多邊形及紡錘形的形態。

Disease
子宮頸癌

02Classification

Organism
Mouse
Sex
Female
Growth properties
Adherent
Biosafety level
1

03Identifiers

Citation
U14(Cytion 型號 305509)
NCBI TaxID
10090
Cellosaurus
CVCL_9U56

04Handling 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
Seeding density
2–4×10⁴/cm²
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.

05Quality 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。

06Donor

Breed / subspecies
庫明