K562-GFP
Cell line · Product number 305948
K-562-GFP cells are a genetically modified variant of the human chronic myelogenous leukemia (CML) cell line K-562, originally derived from the peripheral...
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01General information
- Description
K-562-GFP细胞是人类慢性粒细胞白血病(CML)细胞系K-562的一种基因改造变体,该细胞系最初源自一名处于急变期的成年患者的外周血。 亲本 K-562 细胞系的特征是存在费城染色体,这导致产生具有构成性酪氨酸激酶活性的 BCR-ABL 融合蛋白,从而驱动细胞不受控制地增殖和存活。 K-562 细胞表现出红白血病的特征,在特定的实验条件下可被诱导沿红细胞、巨核细胞或单核细胞谱系分化,使其成为研究造血分化和白血病生物学的多功能模型。
将绿色荧光蛋白(GFP)导入 K-562 细胞,可实现体外和体内白血病细胞行为的实时可视化和追踪。 K-562-GFP细胞广泛应用于涉及细胞增殖、迁移和药物反应的检测,以及在共培养系统中研究其与基质细胞或免疫细胞的相互作用。荧光标记便于进行流式细胞术、活细胞成像和高通量筛选等应用。
基因改造:通过复制缺陷型慢病毒转导进行稳定改造,以表达ZsGreen1绿色荧光蛋白报告基因;在嘌呤霉素(1–5 µg/mL)筛选下维持为多克隆细胞群。S1/BSL-1级生物安全等级。
- Tissue
- 胸腔积液
- Disease
- 慢性髓性白血病
02Classification
- Organism
- Human
- Sex
- Female
- Growth properties
- Suspension
- 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
- 1
03Characteristics
- Morphology
- 淋巴母细胞样
- Cell type
- 淋巴母细胞
04Identifiers
- Citation
- K562-GFP(Cytion 产品编号 305948)
- NCBI TaxID
- 9606
- Cellosaurus
- CVCL_1G55
05Expression and susceptibility
- Antigen expression
- ZsGreen1(绿色荧光蛋白)
- Mutational profile
- 突变:p.Gln136fs*13,纯合子
06Handling 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
- 无
- Subculturing
- 通过定期添加或更换培养基来维持细胞培养。初始培养时细胞密度应为5×10⁵个细胞/毫升,并保持细胞浓度在3×10⁵至1×10⁶个细胞/毫升范围内以获得最佳生长效果。
- Seeding density
- 0.3 至 1 × 10⁶ 个细胞/毫升
- Fluid renewal
- 每周 2 至 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
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
- 53 年
- Ethnicity
- 高加索人
09Legal 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
