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RAW 264.7-GFP
Tissue
腹水
Disease
小鼠白血病
Synonyms
GFP/RAW264.7

Documents

RAW 264.7-GFP

Cell line · Product number 305699

RAW 264.7-GFP cells are a genetically modified derivative of the murine RAW 264.7 macrophage-like cell line, which originates from an adult mouse tumor of...

US$800.00excl. tax and shipping
In stock
Lot
Recommended (freshest lot)
  • CoA
1

01General information

Description

RAW 264.7-GFP 細胞是小鼠 RAW 264.7 巨噬細胞樣細胞系的基因改造衍生物,其源自成年小鼠單核細胞/巨噬細胞系系的腫瘤。 這些細胞經過基因工程改造,可表達綠色螢光蛋白(GFP),從而能在螢光顯微鏡下即時觀察細胞形態、遷移及吞噬活性等細胞過程。 其親本 RAW 264.7 細胞系廣泛用作先天免疫反應的模型,因為它保留了活化巨噬細胞的許多功能特徵,包括在受到刺激時產生細胞因子、一氧化氮和活性氧的能力。

透過整合 GFP,可對體外巨噬細胞的行為進行動態、非侵入性的監測,使 RAW 264.7-GFP 細胞在活細胞成像研究及高內容篩選應用中具有特別重要的價值。 這些細胞對脂多醣(LPS)和干擾素-γ 等促炎性刺激有強烈的反應,從而激活包括 NF-κB 和 MAPK 在內的訊號傳導途徑。這使它們成為研究炎症訊號傳導、宿主-病原體相互作用以及藥理學製劑對巨噬細胞活化的影響的有用模型。 然而,與親本細胞系相同,研究人員應留意,相較於原代巨噬細胞,RAW 264.7 衍生的模型可能在調節路徑上有所改變。

基因改造:透過複製無能的慢病毒轉導進行穩定改造,以表達 ZsGreen1 綠色螢光蛋白報告基因;在嘌呤黴素(1–5 µg/mL)篩選下維持為多克隆細胞群。S1/BSL-1 防護等級。

Tissue
腹水
Disease
小鼠白血病
Synonyms
  • GFP/RAW264.7

02Classification

Organism
Mouse
Sex
Male
Growth properties
Adherent
GMO status
GMO-S1
GMO status details
GMO-S1: This cell line contains a stably integrated ZsGreen1 green fluorescent protein reporter introduced via replication-incompetent lentiviral transduction. The resulting polyclonal cell population was maintained under puromycin selection (1–5 µg/mL). S1 containment is required. This classification applies only within Germany and may differ elsewhere.
Biosafety level
2

03Identifiers

Citation
RAW 264.7-GFP(Cytion 型號 305699)
NCBI TaxID
10090
Cellosaurus
CVCL_D7C8

04Expression and susceptibility

Antigen expression
ZsGreen1(綠色螢光蛋白)

05Handling 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 10% 的胎牛血清(FBS)
Dissociation reagent
Accutase
Seeding density
2 至 5 × 10⁴ 個細胞/cm²
Fluid renewal
每週 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 200 x g for 5 minutes, carefully discard the supernatant containing freezing medium.

  7. Follow the procedure described under Post-Thaw Recovery

06Quality control and cryopreservation

Freeze medium
作為冷凍保存培養基,我們使用完全生長培養基加上 10% 的二甲基亞砜(DMSO),以確保解凍後的細胞存活率達到適當水準。
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
成人