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NCI-H460
Tissue
肺
Disease
肺大細胞癌
Metastatic site
胸腔積液
Synonyms
NCI-H460, NCI.H460, H-460, NCIH460, NCI-HUT-460, NCI-460

Documents

NCI-H460

Cell line · Product number 305020

NCI-H460, also known as H460, was derived from a male patient with large cell lung carcinoma. NCI-H460 cells are adherent cells growing twice as fast than th...

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  • CoA
1

01General information

Description
NCI-H460(亦稱 H460)源自一名患有大細胞肺癌的男性患者。 NCI-H460 細胞為黏附性細胞,其生長速度是 A549 細胞的兩倍,在添加 10% 胎牛血清(FBS)的 RPMI 1640 培養基中,倍增時間為 33 小時。該細胞可在體外及體內模型(包括裸鼠)中形成腫瘤。 研究顯示,NCI-H460 細胞的 p53 mRNA 表達量與正常肺組織相當,且未呈現明顯的 DNA 結構異常。該細胞對角蛋白和維曼汀呈陽性染色,但對神經絲三聚體蛋白呈陰性。 同工酶分析顯示,HPRT 定位於這些非小細胞肺癌細胞系的細胞表面。 AK-1、ES-D 和 Me-2 同工酶的表達水準為 1,而 G6PD、PGM1 和 PGM3 同工酶的表達水準分別為 B 和 1-2。 這些細胞具有低三倍體核型,模態染色體數為 57,範圍介於 53 至 65 之間。 所有細胞均具有七條標記染色體,包括 der(9)t(1;9)(q21;p24)、der(9)t(7;9)(p11;p22)、 t(10q14q)、der(16)t(7;16)(q11.23;q22)。 由於其 p53 mRNA 表現水平極高,因此成為研究非小細胞肺癌分子機制之理想模型。
Tissue
肺
Disease
肺大細胞癌
Metastatic site
胸腔積液
Synonyms
  • NCI-H460
  • NCI.H460
  • H-460
  • NCIH460
  • NCI-HUT-460
  • NCI-460

02Classification

Organism
Human
Sex
Male
Growth properties
Adherent
Biosafety level
1

03Characteristics

Morphology
上皮性
Tumourigenic
是的

04Identifiers

Citation
H-460(Cytion 型號 305020)
NCBI TaxID
9606
Cellosaurus
CVCL_0459

05Handling and culture

Culture medium
RPMI 1640,含 2.0 mM 穩定谷氨醯胺,含 2.0 g/L NaHCO₃(Cytion 產品編號 820700a)
Incubation atmosphere

37°C,5% CO₂,加濕環境。

Supplements
在培養基中加入 10% 的胎牛血清(FBS)
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。

07Donor

Ethnicity
歐洲的