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Detroit-562
Tissue
咽部
Disease
癌
Metastatic site
胸腔積液
Synonyms
DETROIT 562, Detroit 562, Detroit562, DETROIT562, Det 562, Det. 562, Det562, D562

Documents

Detroit-562

Cell line · Product number 300399

Detroit-562 is a human cell line derived from the metastatic site of a pharyngeal carcinoma in an adult male. Established to serve as a model for squamous...

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01General information

Description

Detroit-562 是一種源自一名成年男性咽癌轉移部位的人類細胞系。該細胞系是作為鱗狀細胞癌的模型而建立的,對於研究腫瘤進展與轉移所涉及的生物及分子機制具有特別重要的價值。 底特律-562 細胞呈現上皮細胞形態,當移植至免疫缺陷小鼠體內時,能夠形成鱗狀細胞癌,因此成為癌症研究中一個可靠的體內模型。

該細胞系已被廣泛應用於研究癌症發展中至關重要的細胞訊號傳導路徑,例如涉及表皮生長因子受體(EGFR)的路徑。 研究人員亦利用 Detroit-562 細胞來探索潛在的治療方法,包括藥物篩選及放射治療的療效評估。該細胞系對各種化療藥物的反應性,使其成為評估新型抗癌化合物藥理學特性的關鍵工具。

Tissue
咽部
Disease
癌
Metastatic site
胸腔積液
Synonyms
  • DETROIT 562
  • Detroit 562
  • Detroit562
  • DETROIT562
  • Det 562
  • Det. 562
  • Det562
  • D562

02Classification

Organism
Human
Sex
Female
Growth properties
Adherent
Biosafety level
1

03Characteristics

Morphology
上皮樣

04Identifiers

Citation
底特律-562(Cytion 型號 300399)
NCBI TaxID
9606
Cellosaurus
CVCL_1171

05Expression and susceptibility

Protein expression
P53 陽性
Isoenzymes
G6PD, B
Reverse transcriptase
否
Products
角蛋白

06Handling 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 約需 4 天即可形成密合的細胞層
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.

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
成人
Ethnicity
高加索人