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U14
Disease
宫颈癌

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细胞系是一种小鼠宫颈鳞状细胞癌模型,最初是通过用20-甲基胆蒽处理小鼠子宫颈而建立的。U14细胞已广泛应用于癌症研究,特别是在研究肿瘤侵袭、转移及药物筛选方面。 当移植到小鼠体内时,该细胞系表现出较高的转移潜力,淋巴转移率和肺转移率分别为95%和80%。体外实验中,该细胞表现出上皮细胞特征,包括细胞角蛋白染色阳性,且呈现多边形和梭形形态。

Disease
宫颈癌

02Classification

Organism
Mouse
Sex
Female
Growth properties
Adherent
Biosafety level
1

03Identifiers

Citation
U14(Cytion 产品编号 305509)
NCBI TaxID
10090
Cellosaurus
CVCL_9U56

04Handling and culture

Culture medium
RPMI 1640,w:2.0 mM 稳定谷氨酰胺,w:2.0 g/L NaHCO3(Cytion 文章编号 820700a)。
Incubation atmosphere

37°C, 5%CO2, 加湿环境。

Supplements
在培养基中添加 10% 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

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。

06Donor

Breed / subspecies
库明