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MKN-45
Tissue
胃
Disease
胃腺癌
Metastatic site
肝臟
Synonyms
MKN 45, MKN45

Documents

MKN-45

Cell line · Product number 300489

The MKN-45 cell line is a human gastric cancer cell line, derived from the poorly differentiated adenocarcinoma of the stomach. These cells exhibit charac...

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1

01General information

Description

MKN-45 細胞系是一種人類胃癌細胞系,源自分化程度低的胃腺癌。這些細胞展現出胃癌的典型特徵,包括快速增殖及高度的遺傳不穩定性。 MKN-45 細胞常被用於癌症研究,以探討腫瘤生物學、藥物抗性機制,以及與胃癌進展相關的分子途徑。由於其可透過異種移植方式植入免疫缺陷小鼠體內並形成腫瘤,因此成為體內研究的寶貴模型。

MKN-45 細胞本質上屬於上皮細胞,在培養中以黏附細胞形式生長。它們表達多種與胃癌相關的生物標記物,例如癌胚抗原(CEA)和 E-鈣黏蛋白,因此對於診斷與治療研究具有實用價值。 此外,由於 MKN-45 細胞對治療具有反應性,且能模擬人類胃腫瘤的臨床行為,因此常被用於評估化療藥物及標靶療法。研究人員亦利用此細胞系探索基因改造的影響,並開發旨在改善胃癌患者預後的新治療策略。

Tissue
胃
Disease
胃腺癌
Metastatic site
肝臟
Synonyms
  • MKN 45
  • MKN45

02Classification

Organism
Human
Sex
Female
Growth properties
Adherent and suspension
Biosafety level
1

03Identifiers

Citation
MKN-45(Cytion 型號 300489)
NCBI TaxID
9606
Cellosaurus
CVCL_0434

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 20% 經熱滅活的胎牛血清
Dissociation reagent
Accutase
Subculturing
將懸浮細胞收集至 15 毫升試管中,並用不含鈣和鎂的 PBS 輕柔地洗滌貼壁細胞(T25 培養瓶使用 3-5 毫升,T75 培養瓶使用 5-10 毫升)。 加入 Accutase(T25 培養瓶使用 1–2 毫升,T75 培養瓶使用 2.5 毫升),確保完全覆蓋細胞層。讓細胞於室溫下孵育 10 分鐘。 孵育完成後,將懸浮細胞與貼壁細胞合併並進行離心。離心後,小心地將細胞沉澱重新懸浮,並將細胞懸液轉移至裝有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
62年
Ethnicity
日文