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RKO-E6
Tissue
冒號
Disease
結腸癌
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 細胞是一種源自 RKO 細胞系,並經額外致突變處理而衍生的人類結直腸癌細胞系。這些細胞常被用於癌症研究,特別是針對結直腸癌的研究。 RKO 細胞系的 E6 變異株具有獨特的特徵,有助於探究特定基因操作的影響,並研究大腸直腸癌中腫瘤發生與轉移的分子機制。 RKO-E6 細胞具有若干獨特特徵,包括與細胞週期調控、凋亡及 DNA 修復途徑相關的基因變異。這些變異增強了該細胞系在研究結直腸癌背景下基因沉默或過度表達之生物學效應方面的應用價值。 例如,RKO-E6 細胞已被用於研究腫瘤抑制基因和致癌基因對癌細胞行為的影響,包括增殖、侵襲以及對化療藥物的耐藥性。 此外,RKO-E6 細胞對於旨在了解細胞對環境應激因子(例如氧化應激和 DNA 損傷劑)所作出的反應的研究亦頗具價值,這些應激因子與大腸直腸癌的病理機制及進展密切相關。 其強健的生長特性與遺傳穩定性,使其成為高通量篩選試驗中評估新型抗癌化合物療效的寶貴模型。 總而言之,RKO-E6 細胞為深化我們對大腸直腸癌生物學的認識,以及針對這種常見且往往致命的疾病開發和測試新型治療策略,提供了關鍵的模型。
Tissue
冒號
Disease
結腸癌
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
上皮性

04Identifiers

Citation
RKO-E6(Cytion 型號 305135)
NCBI TaxID
9606
Cellosaurus
CVCL_3787

05Handling and culture

Culture medium
EMEM(MEM Eagle),含 2 mM L-谷氨醯胺,含 2.2 g/L NaHCO₃,含 EBSS(Cytion 產品編號 820100a)
Incubation atmosphere

37°C,5% CO₂,加濕環境。

Supplements
在培養基中加入 10% FBS 和 1% NEAA
Dissociation reagent
Accutase
Subculturing
從貼壁細胞中移除舊培養基,並用不含鈣和鎂的PBS洗滌細胞。T25培養瓶使用3-5毫升PBS,T75培養瓶則使用5-10毫升。 接著,用 Accutase 完全覆蓋細胞,T25 培養瓶使用 1–2 毫升,T75 培養瓶則使用 2.5 毫升。將細胞於室溫下孵育 8–10 分鐘,使其脫附。 孵育後,將細胞與 10 毫升培養基輕柔混合以使其重新懸浮,然後以 300xg 離心 3 分鐘。棄去上清液,將細胞重新懸浮於新鮮培養基中,並轉移至已預先加入新鮮培養基的新培養瓶中。
Fluid renewal
每週 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

透過基於 PCR 的檢測及基於發光原理的支原體檢測方法,排除支原體污染的可能性。

為確保無細菌、真菌或酵母菌污染,每日均會對細胞培養物進行目視檢查。

Freeze medium
作為冷凍保存培養基,我們使用完全生長培養基(含 FBS)+ 10% DMSO 以確保解凍後的存活率,或使用 CM-1 (Cytion 型號 800100),該培養基含有經優化的滲透保護劑和代謝穩定劑,可提升細胞恢復率並減輕冷凍誘導的壓力。
Storage conditions

若需長期保存,請將試管置於溫度約為 −150 至 −196 °C 的氣相液氮中。僅在轉移至液氮之前,可將試管暫時存放於 −80 °C 環境中,作為短期過渡步驟。

Shipping conditions

冷凍保存的細胞系會以乾冰冷藏,並裝於經驗證的隔熱包裝中,包裝內含有足夠的冷媒,以確保在運輸過程中溫度維持在約 −78 °C。收到貨物後,請立即檢查容器,並盡快將試管轉移至適當的儲存場所。