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NCI-H2347
Tissue
肺
Disease
肺腺癌
Synonyms
NCI-H2347, H-2347, NCIH2347

Documents

NCI-H2347

Cell line · Product number 305139

The NCI-H2347 cell line is a human non-small cell lung cancer (NSCLC) cell line derived from a lung adenocarcinoma. This cell line is widely used in studi...

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01General information

Description

NCI-H2347 細胞系是一種源自肺腺癌的人類非小細胞肺癌(NSCLC)細胞系。該細胞系廣泛應用於肺癌生物學研究,特別是涉及腫瘤抑制基因突變,以及與細胞凋亡、化療耐藥性及病毒基癌症療法相關的通路研究。 NCI-H2347 具有野生型 p53,這與許多攜帶 p53 突變的肺癌細胞系形成對比,使其成為研究基於 p53 狀態之治療反應差異的相關模型。

該細胞系已被用於實驗,以測試新型治療方法的療效,例如 ONYX-015——一種經基因改造的腺病毒,能選擇性地在 p53 功能失活的腫瘤細胞內複製並使其裂解。 雖然 ONYX-015 在攜帶 p53 突變的肺癌細胞系(如 NCI-H522)中極為有效,但對攜帶野生型 p53 的 NCI-H2347 而言,其效果則有限。 此外,NCI-H2347 亦曾被用於針對 MET 訊號傳導的研究,特別是與對 EGFR 酪胺酸激酶抑制劑(TKIs)的抗藥性相關的研究。 研究顯示,儘管在此細胞系中未觀察到 MET 基因擴增,但其 MET 蛋白仍可被 EGFR 突變所活化,這表明 MET 與 EGFR 訊號傳導路徑之間存在複雜的相互作用。

Tissue
肺
Disease
肺腺癌
Synonyms
  • NCI-H2347
  • H-2347
  • NCIH2347

02Classification

Organism
Human
Sex
Female
Growth properties
Adherent
Biosafety level
1

03Characteristics

Morphology
上皮性

04Identifiers

Citation
NCI-H2347(Cytion 型號 305139)
NCBI TaxID
9606
Cellosaurus
CVCL_1550

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

Donor age
54年
Ethnicity
歐洲的