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A549-RFP
Tissue
肺部
Disease
肺腺癌
Synonyms
A 549, A549, NCI-A549, A549/ATCC, A549 ATCC, A549ATCC, hA549

Documents

A549-RFP

Cell line · Product number 305659

A549-RFP is a fluorescently labeled derivative of the human A549 lung adenocarcinoma cell line, engineered to constitutively express red fluorescent prote...

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

01General information

Description

A549-RFP 是人 A549 肺腺癌细胞系的荧光标记衍生物,经过基因工程改造,可持续表达红色荧光蛋白(RFP),以便进行实时可视化和追踪。 亲本 A549 细胞系源自一名成年供体的肺腺癌,具有上皮形态和贴壁生长特征。A549 细胞保留了 II 型肺泡上皮细胞的特征,包括细胞角蛋白的表达以及产生表面活性剂相关蛋白的能力。 稳定RFP表达盒的引入使细胞能够持续发出荧光,同时不会显著改变亲本细胞系的固有增殖和代谢特性,因此A549-RFP适用于纵向成像研究。

在大型癌细胞面板中对 A549 细胞进行的功能表征表明,细胞大小、蛋白质含量和蛋白质合成速率与细胞体积呈正相关,且较大的细胞往往增殖较慢。 在比较分析中,A549细胞属于相对较小、增殖较快的上皮癌细胞系,这与那些维明蛋白表达较高、E-钙粘蛋白水平较低、体积较大且更具间质样特征的细胞形成鲜明对比。 这些代谢和表型差异对于实验解释具有重要意义,因为蛋白质合成率和代谢通量与细胞大小成正比,并影响细胞对靶向增殖或 mTOR 调节的合成代谢通路的药物的敏感性。RFP 修饰既保留了 A549 细胞用于此类代谢和药理学研究的适用性,又实现了直接可视化。

A549-RFP 广泛应用于共培养系统、原位和异位异种移植模型以及侵袭或转移实验中,在这些实验中,荧光标记有助于将肿瘤细胞与基质或宿主成分区分开来。 稳定的红色荧光支持多种应用,包括活细胞成像、高通量筛选、基于流式细胞术的定量分析以及体内光学成像。 作为一种特征明确的肺腺癌模型的可追溯变体,A549-RFP 为在体外和体内环境下研究肿瘤细胞增殖、上皮-间质转化、药物反应以及肿瘤-微环境相互作用提供了一个强大的平台。

基因修饰:通过复制缺陷型慢病毒转导进行稳定修饰,以表达红色荧光蛋白(RFP)报告基因;在嘌呤霉素筛选(1–5 µg/mL)条件下维持为多克隆细胞群。S1/BSL-1 生物安全等级。

Tissue
肺部
Disease
肺腺癌
Synonyms
  • A 549
  • A549
  • NCI-A549
  • A549/ATCC
  • A549 ATCC
  • A549ATCC
  • hA549

02Classification

Organism
Human
Sex
Male
Growth properties
Adherent
GMO status
GMO-S1
GMO status details
GMO-S1: This cell line contains a stably integrated red fluorescent protein (RFP) 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
1

03Characteristics

Doubling time
20-40小时

04Identifiers

Citation
A549-RFP(赛通产品编号 305659)
NCBI TaxID
9606
Cellosaurus
CVCL_0023

05Expression and susceptibility

Antigen expression
RFP(红色荧光蛋白)
Mutational profile
突变:p.Gly12Ser,纯合子;突变:p.Gln37Ter,纯合子

06Handling and culture

Culture medium
DMEM:Ham's F12(1:1),含:3.1 g/L 葡萄糖,2.5 mM L-谷氨酰胺,15 mM HEPES,0.5 mM 丙酮酸钠,1.2 g/L NaHCO3(Cytion 产品编号 820400a)
Incubation atmosphere

37°C, 5%CO2, 加湿环境。

Supplements
在培养基中添加 10% FBS
Dissociation reagent
Accutase
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

07Quality 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。

08Donor

Donor age
58 年
Ethnicity
高加索人