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NCI-H1373
Tissue
肺
Disease
腺癌
Synonyms
H1373, H-1373, NCIH1373

Documents

NCI-H1373

Cell line · Product number 305597

The NCI-H1373 cell line is a human non-small cell lung carcinoma (NSCLC) model harboring the KRAS G12C mutation, which is frequently associated with oncog...

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01General information

Description

NCI-H1373 細胞系是一種攜帶 KRAS G12C 突變的人類非小細胞肺癌(NSCLC)模型,該突變常與肺腺癌的致癌過程相關。 此突變會導致下游訊號傳導路徑(例如細胞分裂原活化蛋白激酶(MAPK)和磷脂酰肌醇 3-激酶(PI3K)路徑)持續活化,從而驅動腫瘤增殖與存活。 NCI-H1373 廣泛應用於研究中,用於評估針對 KRAS G12C 及相關致癌途徑的新型治療藥物的療效。

在臨床前研究中,NCI-H1373 已被用於評估新興的 KRAS G12C 抑制劑(如 ASP2453)的活性。該抑制劑展現出對突變型 KRAS 蛋白的強效、選擇性且共價結合能力,並能在體外抑制 KRAS 介導的訊號傳導及腫瘤細胞增殖。 該細胞系亦被應用於異種移植模型中,在該模型中,ASP2453 誘導了顯著的腫瘤退縮。此活性凸顯了 NCI-H1373 在理解新型 KRAS 抑制劑治療潛力與作用機制方面的實用價值。

此外,NCI-H1373 已被應用於全基因組 CRISPR 基因敲除篩選,以探索對 PARP7 抑制劑產生抗藥性的機制。 研究結果顯示,芳香烴受體(AHR)訊號傳導的喪失與對 PARP7 抑制的抗藥性有關,這為潛在的聯合治療策略提供了依據。此類研究證實了該細胞系在識別抗藥性途徑以及優化針對 KRAS G12C 突變非小細胞肺癌(NSCLC)的標靶治療方面的作用。

Tissue
肺
Disease
腺癌
Synonyms
  • H1373
  • H-1373
  • NCIH1373

02Classification

Organism
Human
Sex
Male
Growth properties
Adherent
Biosafety level
1

03Characteristics

Morphology
上皮樣

04Identifiers

Citation
NCI-H1373(Cytion 型號 305597)
NCBI TaxID
9606
Cellosaurus
CVCL_1465

05Expression and susceptibility

Mutational profile
突變:KRAS,p.Gly12Cys (c.34G>T),純合子;突變:TP53,p.Pro47Leu (c.140C>T),純合子

06Handling 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培養瓶需使用3-5毫升PBS,T75培養瓶則需使用5-10毫升。 接著,用 TrypLE Express 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,使其脫落。 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.

07Quality 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。

08Donor

Donor age
56年
Ethnicity
非裔美國人