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HT-3
Tissue
子宫颈
Disease
癌症
Metastatic site
淋巴结
Synonyms
HT3

Documents

HT-3

Cell line · Product number 305521

HT-3 is a human cervical cancer cell line derived from squamous cell carcinoma and is noted for being HPV-negative. This cell line has been widely used in...

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1

01General information

Description

HT-3是一种源自鳞状细胞癌的人宫颈癌细胞系,其特点是HPV阴性。该细胞系已被广泛应用于以细胞生理学为重点的研究中,特别是在离子通道调节、信号转导以及细胞骨架动力学在细胞过程中的作用等方面。 HT-3 细胞表现出体积敏感性氯离子(Cl⁻)通道的激活,这对它们在渗透应激下发生调节性体积减少(RVD)反应至关重要。 该机制通过允许 Cl⁻ 和 K⁺ 离子外流,从而促进水分从细胞内排出,以在细胞肿胀后恢复正常体积,这对维持细胞内稳态至关重要。

对 HT-3 细胞系的研究突显了特定蛋白激酶 C(PKC)同工型在体积调节中的作用。值得注意的是,PKC-α 被认为参与了体积敏感性 Cl⁻ 通道的激活和 RVD 反应。 实验数据表明,无论使用化学阻断剂还是同工型特异性抗体抑制PKC-α,都会显著损害这些通道的功能,导致细胞肿胀时间延长并破坏RVD。这突显了PKC-α作为HT-3细胞对低渗应激反应的调节因子所发挥的作用。

此外,已有研究证实,细胞骨架的完整性(尤其是肌动蛋白丝和微管)对于HT-3细胞中Cl⁻通道的正常激活和有效的体积分调至关重要。 使用细胞松弛素B等药物破坏肌动蛋白丝会增强Cl⁻传导并影响细胞体积恢复,而如鬼笔环肽等稳定剂则会抑制这些通道的活性并延迟RVD。 微管调节化合物(如秋水仙碱和紫杉醇)则产生不同的影响,这凸显了这些癌细胞中细胞骨架结构与离子通道功能之间的复杂相互作用。

Tissue
子宫颈
Disease
癌症
Metastatic site
淋巴结
Synonyms
  • HT3

02Classification

Organism
Human
Sex
Female
Growth properties
Adherent
Biosafety level
1

03Characteristics

Morphology
上皮样
Tumourigenic
是,在裸鼠身上

04Identifiers

Citation
HT-3(Cytion 产品编号 305521)
NCBI TaxID
9606
Cellosaurus
CVCL_1293

05Expression and susceptibility

Antigen expression
表达的基因:A型血,Rh+
Oncogenes
P53+,pRB+
Mutational profile
突变:TP53,p.Gly245Val (c.734G>T)

06Handling and culture

Culture medium
McCoys 5a,w:3.0 克/升葡萄糖,w:稳定谷氨酰胺,w:2.0 毫摩尔丙酮酸钠,w:2.2 克/升 NaHCO3(Cytion 文章编号 820200a)。
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。
Fluid renewal
每周 2 至 3 次
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
58 年
Ethnicity
高加索人