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CDNR4
Tissue
乳房
Disease
腺癌

Documents

CDNR4

Cell line · Product number 400391

The CDNR4 cell line comprises a specialized subset originating from the COMMA-D cell line, known for modeling mouse mammary carcinoma. This clonal subpopu...

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01General information

Description

CDNR4 細胞系是由 COMMA-D 細胞系衍生而來的一種特殊亞群,該細胞系以能模擬小鼠乳腺癌而聞名。這克隆亞群已經過廣泛的特性分析,揭示出一系列獨特的特性與功能。 CDNR4 細胞最引人注目的特徵之一,是其與乳腺幹細胞的相似性,這使其成為探索幹細胞生物學、致癌機制以及細胞群體內異質性等領域的重要資源。 這些細胞是透過轉染攜帶卡那黴素和新黴素抗性基因(Tn5 基因)的轉座子而開發出來的,此過程促使各種引人入勝的特徵與能力浮現,包括其分化為癌前及腫瘤表型的潛力。

CDNR4 源自 COMMA-D 細胞系,該細胞系最初是透過相位對比顯微鏡、免疫細胞化學染色、DNA 含量分析以及致癌潛力評估等多種技術,針對其細胞異質性進行研究,而 CDNR4 則作為一個獨特的克隆而脫穎而出。 透過特定的轉染與篩選方法,成功分離出如 CDNR4 般的克隆亞群,每個亞群均保留了原始 COMMA-D 親代細胞所呈現的某種程度的異質性。這種異質性的保留凸顯了這些細胞群體的複雜本質,並提升了 CDNR4 細胞在細胞分化與癌症進展相關研究中的價值。

Tissue
乳房
Disease
腺癌

02Classification

Organism
Mouse
Sex
Female
Growth properties
Adherent
Biosafety level
1

03Identifiers

Citation
CDNR4(Cytion 型號 400391)
NCBI TaxID
10090
Cellosaurus
CVCL_5719

04Handling and culture

Culture medium
DMEM,含 4.5 g/L 葡萄糖、4 mM L-谷氨醯胺、3.7 g/L NaHCO₃ 及 1.0 mM 丙酮酸鈉(Cytion 產品編號 820300a)
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。
Seeding density
建議為 2 × 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.

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
1 年