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SiHa
Tissue
子宫颈
Disease
与人类乳头瘤病毒有关的宫颈鳞状细胞癌
Synonyms
Siha, SIHA

Documents

SiHa

Cell line · Product number 305023

SiHa cells are a human cervical squamous cell carcinoma cell line that has been widely used in research for several decades. They were isolated from prima...

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1

01General information

Description

SiHa 细胞是一种人宫颈鳞状细胞癌细胞系,几十年来一直被广泛用于研究。它们是从一名 55 岁的日本女性鳞状细胞癌患者的原发性子宫活检片段中分离出来的。由于其独特的遗传特性,研究宫颈癌和其他相关疾病的研究人员对这种细胞系非常感兴趣。

研究发现,SiHa 细胞表达 p53+ 和 pRB+ 基因,这些基因参与细胞周期调控、DNA 修复和肿瘤抑制。这些基因使 SiHa 细胞成为研究癌症发生和发展分子机制的理想模型。此外,SiHa 细胞还是一种合适的转染宿主,使其成为基因表达研究的绝佳工具。

SiHa 细胞具有高倍核型,平均染色体数在 69 到 72 之间。SiHa 细胞是 HPV-16 阳性细胞,每个细胞整合 1 到 2 个病毒基因组拷贝。细胞具有致瘤性,可在裸鼠体内形成分化不良的表皮样癌(III 级)。这使它们成为研究癌症进展和测试抗癌药物的极佳模型。

SiHa 细胞系表达多种同工酶,包括 AK-1、ES-D、G6PD、GLO-I、Me-2、PGM1 和 PGM3。电子显微镜显示细胞质中有丰富的碳丝(tonofilaments),细胞连接处有脱膜小体(desmosomes)。SiHa 细胞具有粘附生长特性,在 10% FBS 培养基中的倍增时间为 17 小时,在 5% FBS 培养基中的倍增时间为 21 小时。92%的 SiHa 细胞都有上皮细胞粘附分子(EpCAM)表达,这表明它们来源于上皮细胞。它们有很强的细胞角蛋白表达,但没有波形蛋白表达。

Tissue
子宫颈
Disease
与人类乳头瘤病毒有关的宫颈鳞状细胞癌
Synonyms
  • Siha
  • SIHA

02Classification

Organism
Human
Sex
Female
Growth properties
Adherent
Biosafety level
1

03Characteristics

Morphology
上皮细胞
Tumourigenic
是

04Identifiers

Citation
SiHa(Cytion 产品编号 305023)
NCBI TaxID
9606
Cellosaurus
CVCL_0032

05Handling and culture

Culture medium
EMEM(MEM Eagle),w:2 mM L-谷氨酰胺,w:2.2 g/L NaHCO3,w:EBSS(Cytion 文章编号 820100a)。
Incubation atmosphere

37°C, 5%CO2, 加湿环境。

Supplements
在培养基中添加 10% FBS 和 1% NEAA
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.

06Quality 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。

07Donor

Donor age
55 岁
Ethnicity
亚洲