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NCI-H196
Tissue
肺
Disease
肺小細胞癌
Metastatic site
胸腔積液
Synonyms
NCI-H196, H-196, NCIH196

Documents

NCI-H196

Cell line · Product number 300390

NCI-H196 is a small-cell lung cancer (SCLC) cell line used to study the mechanisms of cancer progression, chemotherapy resistance, and cellular responses ...

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01General information

Description

NCI-H196 是一種小細胞肺癌(SCLC)細胞系,常用於研究癌症進展、化療抗藥性以及細胞對氧化應激的反應機制。 針對 NCI-H196 的研究顯示,該細胞系對促氧化劑吡咯烷二硫代氨基甲酸酯(PDTC)的細胞毒性作用具有敏感性。PDTC 會誘導 S 期細胞週期停滯,並以劑量依賴性方式顯著降低 NCI-H196 細胞的存活率。 這種細胞毒性被歸因於氧化應激的誘導,其證據在於活性氧(ROS)的增加以及氧化應激相關基因表達的變化。 添加 N-乙醯-L-半胱胺酸(NAC)等抗氧化劑可有效逆轉 PDTC 誘導的細胞毒性,這證實了氧化壓力在細胞死亡中的作用。

進一步研究顯示,PDTC 可增強順鉑(一種用於治療小細胞肺癌的一線化療藥物)的細胞毒性。將低劑量順鉑與無毒濃度的 PDTC 聯合使用,可在 NCI-H196 細胞中產生協同細胞毒性作用。 此聯合療法之所以有效,被認為是因 PDTC 可下調 ATP7A 的表達——ATP7A 是一種與順鉑抗藥性相關的銅外排轉運蛋白。透過抑制 ATP7A,PDTC 可能增加細胞內銅濃度,並增強 NCI-H196 細胞對順鉑的敏感性,凸顯其作為小細胞肺癌輔助療法的潛力。

Tissue
肺
Disease
肺小細胞癌
Metastatic site
胸腔積液
Applications
3D 細胞培養、癌症研究
Synonyms
  • NCI-H196
  • H-196
  • NCIH196

02Classification

Organism
Human
Sex
Male
Growth properties
Adherent
Biosafety level
1

03Identifiers

Citation
NCI-H196(Cytion 型號 300390)
NCBI TaxID
9606
Cellosaurus
CVCL_1509

04Handling 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。
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.

05Quality 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。

06Donor

Donor age
68年
Ethnicity
歐洲的