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K562-GFP
Tissue
胸腔积液
Disease
慢性髓性白血病

Documents

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
  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
53 年
Ethnicity
高加索人