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EBC-1
Tissue
肺
Disease
鱗狀細胞癌
Metastatic site
皮膚
Synonyms
EBC-1/original, EBC1

Documents

EBC-1

Cell line · Product number 305539

EBC-1 is a human lung squamous cell carcinoma cell line, primarily noted for its relevance in studying mechanisms related to lung cancer, particularly non...

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01General information

Description

EBC-1 是一種人類肺鱗狀細胞癌細胞系,主要因其對研究肺癌(特別是非小細胞肺癌,NSCLC)相關機制的重要性而備受矚目。此細胞系的特徵在於 MET 基因擴增,該基因已被證實與驅動腫瘤生長及治療抗藥性的致癌訊號傳導路徑有關。 MET受體酪胺酸激酶的活化(通常由肝細胞生長因子(HGF)誘導)在這些細胞的增殖、存活及轉移過程中扮演重要角色。MET訊號傳導的異常是EBC-1具有侵襲性腫瘤特徵的關鍵因素,使其成為研究針對MET抑制的標靶治療的重要模型。

利用 EBC-1 細胞進行的研究,已探討了針對克唑替尼等 MET 抑制劑的各種抗藥性機制。該細胞系已顯示出透過 PAI-1 上調及上皮-間質轉化(EMT)等途徑所產生的獲得性抗藥性,這為治療帶來了挑戰。 此外,研究顯示丁酸鈉能調控 EBC-1 細胞的基因表達,這表明組蛋白去乙醯酶抑制劑在影響基因轉錄方面具有潛在應用價值。這些發現凸顯了 EBC-1 在治療抗藥性研究以及針對 MET 擴增型肺癌開發新型治療策略方面的關鍵重要性。

Tissue
肺
Disease
鱗狀細胞癌
Metastatic site
皮膚
Synonyms
  • EBC-1/original
  • EBC1

02Classification

Organism
Human
Sex
Male
Growth properties
Adherent
Biosafety level
1

03Identifiers

Citation
EBC-1(Cytion 型號 305539)
NCBI TaxID
9606
Cellosaurus
CVCL_2891

04Expression and susceptibility

Mutational profile
突變:DDR2,p.Thr681Ile (c.2042C>T),雜合子;突變:EGFR,p.Leu858Arg (c.2573T>G),雜合子; 突變:TP53,p.Glu171Ter (c.511G>T),純合子

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。
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。

07Donor

Donor age
69年
Ethnicity
台灣人