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...
Recommended (freshest lot)
01General information
- Description
RAW 264.7-GFP细胞是小鼠RAW 264.7巨噬细胞样细胞系的基因改造衍生物,该细胞系源自成年小鼠单核细胞/巨噬细胞谱系的肿瘤。 这些细胞经过基因工程改造,可表达绿色荧光蛋白(GFP),从而能够通过荧光显微镜实时观察细胞形态、迁移和吞噬活性等细胞过程。 亲本 RAW 264.7 细胞系被广泛用作先天免疫反应的模型,因为它保留了活化巨噬细胞的许多功能特征,包括在受到刺激时产生细胞因子、一氧化氮和活性氧的能力。
GFP的引入使得在体外对巨噬细胞的行为进行动态、无创的监测成为可能,这使得RAW 264.7-GFP细胞在活细胞成像研究和高通量筛选应用中具有特别重要的价值。 这些细胞对脂多糖 (LPS) 和干扰素-γ 等促炎性刺激反应强烈,从而激活包括 NF-κB 和 MAPK 在内的信号传导通路。这使得它们成为研究炎症信号传导、宿主-病原体相互作用以及药理学制剂对巨噬细胞活化的影响的有用模型。 然而,与亲本细胞系一样,研究人员应考虑到,与原代巨噬细胞相比,RAW 264.7衍生的模型在调控通路方面可能存在差异。
基因修饰:通过复制缺陷型慢病毒转导进行稳定修饰,以表达ZsGreen1绿色荧光蛋白报告基因;在嘌呤霉素(1–5 µg/mL)筛选下作为多克隆细胞群维持。S1/BSL-1级生物安全等级。
- 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(赛通产品编号 305699)
- NCBI TaxID
- 10090
- Cellosaurus
- CVCL_D7C8
04Expression and susceptibility
- Antigen expression
- ZsGreen1(绿色荧光蛋白)
05Handling and culture
- Culture medium
- RPMI 1640,w:2.0 mM 稳定谷氨酰胺,w:2.0 g/L NaHCO3(Cytion 文章编号 820700a)。
- Incubation atmosphere
37°C, 5%CO2, 加湿环境。
- Supplements
- 在培养基中添加 10% FBS
- Dissociation reagent
- Accutase
- Seeding density
- 2至5×10⁴个细胞/平方厘米
- Fluid renewal
- 每周 3 次
- Thawing and culturing
Confirm that the vial remains deeply frozen upon delivery, as cells are shipped on dry ice to maintain optimal temperatures during transit.
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.
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.
Perform all subsequent steps under sterile conditions in a flow hood, disinfecting the cryovial with 70% ethanol before opening.
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.
Centrifuge the mixture at 200 x g for 5 minutes, carefully discard the supernatant containing freezing medium.
Follow the procedure described under Post-Thaw Recovery
06Quality control and cryopreservation
- Freeze medium
- 作为冷冻保存培养基,我们使用完全生长培养基+10% DMSO,以获得足够的解冻后存活率。
- Storage conditions
如需长期保存,可将样品瓶放入气相液氮中,温度约为 -150 至 -196 ℃。在-80 °C下保存只能作为转入液氮前的短暂过渡。
- Shipping conditions
冷冻保存的细胞系用干冰装运,并用经过验证的隔热包装和足够的制冷剂在整个运输过程中保持约 -78 °C 的温度。收到货后,应立即检查容器,并立即将小瓶转移到适当的储藏室。
07Donor
- Donor age
- 成人
08Legal and safety
- Safety precautions
- 计划将低温样品储存在液氮中以便将来解冻时,必须遵守严格的安全措施。在将冷冻样本移入或移出液氮罐时,必须穿戴适当的防护手套和防护服,并使用面罩或安全护目镜。这样做是为了降低在移出时因冷冻样品爆炸而造成伤害的风险,因为爆炸可能导致锋利的碎片飞溅。
- Warranty
在Cytion,我们保证产品的质量和活力。 我们保证,从我们的工厂发货后 60 天内,我们的细胞在开始培养时都是存活的。 我们保证所有材料均符合分析证书和/或产品说明书中描述的规格。
有关详细信息,请参阅我们的 <a href="" /Who-we-are/Warranty/"" hreflang=""x-default"">保修声明 。
- Material Transfer Agreement required
- Yes
