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CERV-196
Tissue
子宫颈
Disease
鳞状细胞癌
Synonyms
Cerv-196, MRI-H-196, MRI-H196

Documents

CERV-196

Cell line · Product number 300291

The MRI-H196 cell line, derived from HPV16-positive cervical carcinoma, displays a unique HPV16 transcript expression profile characterized by the presenc...

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01General information

Description

MRI-H196 细胞系来源于 HPV16 阳性宫颈癌,显示出独特的 HPV16 转录本表达谱,其特点是存在全长 L1 转录本,而明显缺乏 E5 全长 RNA。这种模式表明,HPV16 基因组在该细胞系中发生了整合,特别是影响了 E2 区域,并导致 L1 DNA 序列重排。E5 全长 RNA 表达的缺失表明全长早期 RNA 的转录发生了中断,这种中断通常发生在位于 E5 开放阅读框(ORF)下游的多聚腺苷酸化信号处。这种中断表明了 HPV16 基因组的整合状态,其中关键的 E2 区--病毒复制和转录调控的关键--往往在整合到宿主基因组的过程中受损。这种破坏可能会影响包括 E5 在内的下游基因的表达。

MRI-H196 细胞内的这种整合现象凸显了整合后 HPV16 基因组行为的复杂性,强调了该细胞系在研究宫颈癌中与 HPV 整合相关的基因组和转录错综复杂性方面的实用性。了解这些动态变化对于深入了解肿瘤发生机制和 HPV 相关癌症的进展至关重要,这使得 MRI-H196 细胞系成为医学和生物学研究的宝贵资源。

Tissue
子宫颈
Disease
鳞状细胞癌
Synonyms
  • Cerv-196
  • MRI-H-196
  • MRI-H196

02Classification

Organism
Human
Sex
Female
Growth properties
Adherent
Biosafety level
2

03Characteristics

Morphology
上皮样
Tumourigenic
是,在裸鼠身上

04Identifiers

Citation
CERV-196(Cytion 目录号 300291)
NCBI TaxID
9606
Cellosaurus
CVCL_5721

05Expression and susceptibility

Viruses
HPV-16 阳性
Products
细胞角蛋白 8、18、波形蛋白

06Handling and culture

Culture medium
DMEM:Ham's F12(1:1),w: 3.1 g/L 葡萄糖,w: 2.5 mM L-谷氨酰胺,w: 15 mM HEPES,w: 0.5 mM 丙酮酸钠,w: 1.2 g/L NaHCO3(Cytion 文章编号 820400a)。
Incubation atmosphere

37°C, 5%CO2, 加湿环境。

Supplements
在培养基中添加 10% FBS
Dissociation reagent
Accutase
Subculturing
去除粘附细胞上的旧培养基,用不含钙和镁的 PBS 冲洗。T25 烧瓶用 3-5 毫升 PBS,T75 烧瓶用 5-10 毫升。然后用 Accutase 完全覆盖细胞,T25 烧瓶用 1-2 毫升,T75 烧瓶用 2.5 毫升。让细胞在室温下孵育 8-10 分钟,使其分离。孵育后,用 10 毫升培养基轻轻混合细胞使其重悬,然后用 300xg 离心 3 分钟。弃去上清液,用新鲜培养基重悬细胞,并将其转移到已装有新鲜培养基的新烧瓶中。
Seeding density
建议使用1×10⁴个细胞/平方厘米
Fluid renewal
每周 2 至 3 次
Post-thaw recovery
解冻后,将细胞以5×10⁴个细胞/平方厘米的密度接种于培养皿中,使细胞从冷冻过程中恢复并至少贴壁24小时。
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

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。

08Donor

Donor age
49 岁
Ethnicity
非洲