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KHOS-240S
Tissue
骨頭
Disease
骨肉瘤
Synonyms
KHOS240S

Documents

KHOS-240S

Cell line · Product number 300433

KHOS-240S is an osteosarcoma cell line that is derived from human bone sarcoma tissue. This cell line, along with its variants, has been extensively used ...

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01General information

Description

KHOS-240S 是一種源自人類骨肉瘤組織的骨肉瘤細胞系。該細胞系及其變異株已被廣泛應用於骨肉瘤相關研究;骨肉瘤是一種主要影響兒童及年輕成人的原發性惡性骨腫瘤。 骨肉瘤的特徵在於惡性細胞會產生未成熟的骨組織(骨樣組織),且以其侵襲性強及易於早期轉移(特別是轉移至肺部)而聞名。

KHOS-240S 細胞系對多種激酶抑制劑具有抗藥性,包括針對 PI3K-Akt-mTOR 路徑的抑制劑。這種對常見治療標靶的抗藥性,使 KHOS-240S 在研究骨肉瘤的藥物抗藥性機制及探索替代治療策略方面具有特別重要的價值。 研究人員已利用此細胞系篩選多種抗腫瘤藥物及試驗性藥物,並藉此鑑定出可能克服抗藥性機制的化合物。 在利用 KHOS-240S 進行的研究中,與藥物抗藥性及骨肉瘤生物學相關的基因(例如參與 mTOR 訊號傳導路徑的基因)之表達譜,尤為值得關注。

此外,KHOS-240S 亦被用於探索微RNA 的表達模式,這些模式可能與藥物敏感性或抗藥性相關。 該細胞系對 PI3K-Akt-mTOR 路徑抑制劑的特定抗藥性,為理解骨肉瘤如何規避標靶治療提供了關鍵模型,並為開發能提升抗藥性骨肉瘤亞型治療成效的新型治療方法奠定了基礎。

Tissue
骨頭
Disease
骨肉瘤
Synonyms
  • KHOS240S

02Classification

Organism
Human
Sex
Female
Growth properties
Adherent
Biosafety level
1

03Characteristics

Morphology
成纖維細胞樣
Tumourigenic
不

04Identifiers

Citation
KHOS-240S(Cytion 型號 300433)
NCBI TaxID
9606
Cellosaurus
CVCL_2544

05Handling and culture

Culture medium
EMEM(MEM Eagle),含 2 mM L-谷氨醯胺,含 2.2 g/L NaHCO₃,含 EBSS(Cytion 產品編號 820100a)
Incubation atmosphere

37°C,5% CO₂,加濕環境。

Supplements
在培養基中加入 10% FBS 和 1% NEAA
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。
Seeding density
1 × 10⁴ 個細胞/cm²
Fluid renewal
每週 2 至 3 次
Post-thaw recovery
解凍後,將細胞以 5 × 10⁴ 個細胞/cm² 的密度接種於培養皿中,並讓細胞從冷凍過程中恢復,並附著至少 24 小時。
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
13年
Ethnicity
高加索人