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RKO-E6
Tissue
结肠
Disease
结肠癌
Synonyms
RKOE6

Documents

RKO-E6

Cell line · Product number 305135

RKO-E6 cells are a human colorectal carcinoma cell line derived from the RKO cell line through additional mutagenesis. These cells are commonly used in cance...

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01General information

Description
RKO-E6 细胞是一种人类结直肠癌细胞系,由 RKO 细胞系通过额外的诱变得到。这些细胞常用于癌症研究,尤其是结直肠癌研究。RKO 细胞系的 E6 变体具有独特的特征,可用于检测特定基因操作的效果以及研究结直肠癌肿瘤发生和转移的分子机制。 RKO-E6 细胞具有几个独特的特征,包括细胞周期调控、细胞凋亡和 DNA 修复途径相关基因的改变。这些改变增强了该细胞系在研究结直肠癌基因沉默或过表达的生物效应方面的实用性。例如,RKO-E6 细胞已被用于研究肿瘤抑制基因和癌基因对癌细胞行为的影响,包括增殖、侵袭和对化疗药物的抗性。 此外,RKO-E6 细胞还可用于研究细胞对氧化应激和 DNA 损伤剂等环境应激源的反应,这些应激源与结直肠癌的发病机制和进展有关。总之,RKO-E6 细胞为增进我们对结直肠癌生物学的了解,以及开发和测试针对这种流行且往往致命的疾病的新型治疗策略提供了一个重要的模型。
Tissue
结肠
Disease
结肠癌
Synonyms
  • RKOE6

02Classification

Organism
Human
Growth properties
Adherent
GMO status
GMO-S1
GMO status details
GMO-S1: This human colorectal carcinoma cell line (RKO-E6) contains a plasmid encoding HPV-16 E6 under CMV promoter control, possibly including CMV and HPV-6 sequences, enabling E6-dependent transformation studies. The construct is stably integrated. This classification applies only within Germany and may differ elsewhere.
Biosafety level
2

03Characteristics

Morphology
上皮细胞

04Identifiers

Citation
RKO-E6(Cytion 产品编号 305135)
NCBI TaxID
9606
Cellosaurus
CVCL_3787

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。