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HK-ZFN-AURKB-mEGFP 细胞
Tissue
宫颈内口
Disease
腺癌
Metastatic site
原发肿瘤部位(宫颈管)

Documents

HK-ZFN-AURKB-mEGFP 细胞

Cell line · Product number 300173

The HK-ZFN-AURKB-mEGFP cell line is a genetically engineered human cell model designed to express the AURKB (Aurora Kinase B) protein fused with mEGFP (mo...

US$800.00excl. tax and shipping
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1

01General information

Description

HK-ZFN-AURKB-mEGFP 细胞系是一种基因工程人体细胞模型,利用锌指核酸酶(ZFN)技术表达与 mEGFP(单体增强型绿色荧光蛋白)融合的 AURKB(极光激酶 B)蛋白。AURKB 是一种丝氨酸/苏氨酸激酶,在有丝分裂染色体分离、细胞分裂和有丝分裂纺锤体检查点的调控中起着至关重要的作用。通过与 mEGFP 融合,可以实时观察 AURKB 在细胞内的活性和定位,便于详细研究其在细胞分裂过程中的动态行为。

该细胞系是研究有丝分裂分子机制和 AURKB 特定功能的有力工具。mEGFP 的加入实现了基于荧光的检测和活细胞成像,为研究 AURKB 的时空分布提供了洞察力。ZFN 技术的使用确保了精确的基因组整合,保持了 AURKB 表达的真实性。这种模型在癌症研究中尤其有价值,因为 AURKB 通常会过度表达并与肿瘤发生有关,这使它成为治疗干预的潜在靶点。

Tissue
宫颈内口
Disease
腺癌
Metastatic site
原发肿瘤部位(宫颈管)
Applications
Aurora B 激酶(AURKB)生物学;有丝分裂激酶成像;染色体分离;纺锤体检查点;活细胞成像;ZFN 基因组编辑验证;癌症生物学

02Classification

Organism
Human
Sex
Female
Growth properties
Adherent
GMO status
GMO-S1
GMO status details
GMO-S1: This HeLa Kyoto line contains a ZFN-integrated mEGFP fusion at the endogenous AURKB locus for mitotic kinase imaging. This classification applies only within Germany and may differ elsewhere.
Biosafety level
1

03Characteristics

Morphology
具有镶嵌石形状的上皮样细胞
Cell type
上皮细胞

04Identifiers

Citation
HK-ZFN-AURKB-mEGFP(Cytion 目录号 300173)
NCBI TaxID
9606
Cellosaurus
CVCL_VL13
Depositor
艾伦伯格实验室(EMBL)

05Expression and susceptibility

Products
EGFP(增强型绿色荧光蛋白)

06Handling and culture

Culture medium
DMEM,w:4.5 克/升葡萄糖,w:4 毫摩尔 L-谷氨酰胺,w:3.7 克/升 NaHCO3,w:1.0 毫摩尔丙酮酸钠(Cytion 编号 820300a)。
Incubation atmosphere

37°C, 5%CO2, 加湿环境。

Supplements
在培养基中添加 10% FBS
Dissociation reagent
Accutase
Subculturing
去除粘附细胞上的旧培养基,用不含钙和镁的 PBS 冲洗。T25 烧瓶用 3-5 毫升 PBS,T75 烧瓶用 5-10 毫升。然后用 Accutase 完全覆盖细胞,T25 烧瓶用 1-2 毫升,T75 烧瓶用 2.5 毫升。让细胞在室温下孵育 8-10 分钟,使其分离。孵育后,用 10 毫升培养基轻轻混合细胞使其重悬,然后用 300xg 离心 3 分钟。弃去上清液,用新鲜培养基重悬细胞,并将其转移到已装有新鲜培养基的新烧瓶中。
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 300 x g for 3 minutes to separate the cells and carefully discard the supernatant containing residual freezing medium.

  7. Gently resuspend the cell pellet in 10 ml of fresh culture medium. For adherent cells, divide the suspension between two T25 culture flasks; for suspension cultures, transfer all the medium into one T25 flask to promote effective cell interaction and growth.

  8. Adhere to established subculture protocols for continued growth and maintenance of the cell line, ensuring reliable experimental outcomes.

07Quality control and cryopreservation

Sterility

使用基于 PCR 的检测方法和基于发光的支原体检测方法排除支原体污染。

为确保没有细菌、真菌或酵母菌污染,每天都要对细胞培养物进行目视检查。

Freeze medium
我们使用完全生长培养基(包括 FBS)+10% DMSO 或 CM-1(Cytion 商品目录编号 800100)作为冷冻保存培养基,以获得足够的解冻后存活率,CM-1(Cytion 商品目录编号 800100)含有优化的渗透保护剂和代谢稳定剂,可提高恢复能力并减少冷冻引起的应激。
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
30年
Ethnicity
非洲裔美国人