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KYSE520
Tissue
食道
Disease
鳞状细胞癌
Synonyms
KYSE 520, KYSE-520, Kyse520, KYSE0520

Documents

KYSE520

Cell line · Product number 305449

The KYSE520 cell line is a human esophageal squamous cell carcinoma (ESCC) model derived from a primary tumor. It is moderately differentiated and has bee...

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1

01General information

Description

KYSE520 细胞系是一种人类食管鳞状细胞癌 (ESCC) 模型,来源于原发肿瘤。它属于中度分化型,在研究食管癌的上皮-间质可塑性(EMP)方面发挥了重要作用。KYSE520 细胞具有异质性,由上皮样(CD44v+)和间质样(CD44v-)亚群组成。这两个亚群能够相互转化,反映了一个动态的 EMP 过程。这一特性使KYSE520成为研究ESCC中癌症干细胞特征和化疗抗性机制的绝佳模型。

在遗传学上,KYSE520细胞显示出显著的表观遗传调控。在这些细胞中,肿瘤抑制因子JAM3基因的启动子区域未被甲基化,允许其表达。JAM3 通过 Wnt/β-catenin 信号在调节细胞增殖、迁移和侵袭方面发挥作用。维持 KYSE520 中 JAM3 的表达与抑制侵袭性癌症表型有关。

在治疗研究中,KYSE520 细胞被用来探索成纤维细胞生长因子受体样 1(FGFRL1)的作用。研究表明,FGFRL1缺陷的KYSE520细胞表现出肿瘤生长和运动减弱,同时基质金属蛋白酶-1(MMP-1)和成纤维细胞生长因子结合蛋白1(FGFBP1)表达减少。这些发现强调了 FGFRL1 在肿瘤发生中的重要性,并提出了潜在的治疗靶点。此外,KYSE520细胞中的EMP动态和相关分子通路有助于深入了解ESCC的进展和耐药机制,从而促进靶向治疗的开发。

Tissue
食道
Disease
鳞状细胞癌
Synonyms
  • KYSE 520
  • KYSE-520
  • Kyse520
  • KYSE0520

02Classification

Organism
Human
Sex
Female
Growth properties
Adherent
Biosafety level
1

03Characteristics

Morphology
上皮样

04Identifiers

Citation
KYSE520(Cytion 目录号 305449)
NCBI TaxID
9606
Cellosaurus
CVCL_1355

05Expression and susceptibility

Oncogenes
TP53、MYC
Mutational profile
突变:TP53,c.376-2A>T,剪接受体突变

06Handling and culture

Culture medium
Ham's F12,w: 1.0 mM 稳定谷氨酰胺,w: 1.0 mM 丙酮酸钠,w: 1.1 g/L NaHCO3(Cytion 产品编号 820600a)+ RPMI 1640,w: 2.0 mM 稳定谷氨酰胺,w: 2.0 g/L NaHCO3(Cytion 产品编号 820700a);1:1 混合物
Incubation atmosphere

37°C, 5%CO2, 加湿环境。

Supplements
在培养基中添加 2% 的 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
0.6 - 1.2 × 10⁴ 个细胞/平方厘米
Fluid renewal
每周 2 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
日语