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Daoy
Tissue
大腦、小腦
Disease
髓母細胞瘤
Synonyms
DAOY, D324 Med, D-324 Med, D324 MED, D-324MED, D324

Documents

Daoy

Cell line · Product number 305053

The Daoy cell line, established in 1985 by P.F. Jacobsen at the Royal Perth Hospital in Western Australia, is a human cell line derived from a medulloblas...

US$395.00excl. tax and shipping
In stock
Lot
Recommended (freshest lot)
  • CoA
1

01General information

Description

Daoy 細胞系由 P.F. Jacobsen 於 1985 年在西澳的皇家珀斯醫院建立,是一種源自髓母細胞瘤的人類細胞系,而髓母細胞瘤是一種主要發生於兒童的腦腫瘤。 該細胞系源自一名4歲男童後顱窩腫瘤的活體切片。髓母細胞瘤通常位於小腦——這是大腦中對運動控制與協調至關重要的區域——且是兒童中最常見的惡性腦腫瘤。

Daoy 細胞廣泛用作研究髓母細胞瘤生物學的模型系統,涵蓋腫瘤起始、進展及對治療的反應等領域。該細胞系在髓母細胞瘤研究中發揮了關鍵作用,特別是在釐清該疾病的分子與遺傳基礎,以及測試化療藥物方面。 這些細胞展現出惡性髓母細胞瘤的典型特徵,包括快速增殖率,以及移植至免疫缺陷小鼠體內後能形成腫瘤的能力。利用 Daoy 細胞系進行的研究,已為髓母細胞瘤潛在的新療法及治療靶點的開發做出貢獻。

Tissue
大腦、小腦
Disease
髓母細胞瘤
Synonyms
  • DAOY
  • D324 Med
  • D-324 Med
  • D324 MED
  • D-324MED
  • D324

02Classification

Organism
Human
Sex
Male
Growth properties
Adherent
Biosafety level
1

03Characteristics

Morphology
多邊形
Doubling time
34 小時

04Identifiers

Citation
Daoy(Cytion 型號 305053)
NCBI TaxID
9606
Cellosaurus
CVCL_1167

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。
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.

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
4年
Ethnicity
歐洲的