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U-2 OS-ZFN-SNAP-Nup107
Tissue
骨质
Disease
骨肉瘤

Documents

U-2 OS-ZFN-SNAP-Nup107

Cell line · Product number 300294

The U-2 OS-ZFN-SNAP-Nup107 cell line represents a specialized variant of the widely used human osteosarcoma cell line U-2 OS. This particular model has be...

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01General information

Description

U-2 OS-ZFN-SNAP-Nup107 细胞系是广泛使用的人类骨肉瘤细胞系 U-2 OS 的一种特殊变体。这种特殊的细胞株被设计为表达带有 SNAP 标记的核疏导蛋白 Nup107,Nup107 是核孔复合体的关键组成部分,对细胞核和细胞质之间的分子运输至关重要。SNAP 标记技术的集成允许对活体或固定细胞中的 Nup107 进行生物标记和可视化,为研究生理条件下的核胞质转运和核孔复合体结构提供了一个强大的工具。

U-2 OS背景具有多种优势,包括生长速度快和核型特征良好,可支持高通量筛选应用和基因组研究。该细胞系采用的 ZFN(锌指核酸酶)技术有助于进行有针对性的基因组编辑,提高了研究人员研究核转运和其他细胞过程的基因贡献的精确度。这种细胞系特别适用于旨在阐明癌症生物学和细胞生理学中核孔复合体的动态和调控的研究。

由于 U-2 OS-ZFN-SNAP-Nup107 no.294 细胞系的特殊性,它非常适合采用先进的成像技术,包括超分辨率显微镜,以前所未有的细节探索核孔复合体的功能。它也是开发针对与癌症等各种疾病有关的核转运途径的治疗策略的宝贵资源。SNAP 标签组件为进一步的生化和蛋白质组分析增添了多功能性,使其成为细胞和分子生物学领域不可或缺的工具。

Tissue
骨质
Disease
骨肉瘤

02Classification

Organism
Human
Sex
Female
Growth properties
Adherent
GMO status
GMO-S1
GMO status details
GMO-S1: This human osteosarcoma cell line (U2OS-ZFN-SNAP-Nup107 no.294) contains a ZFN-mediated SNAP-Nup107 gene fusion, supporting selective labeling of the Nup107 nuclear pore subunit. The modification is stably present. This classification applies only within Germany and may differ elsewhere.
Biosafety level
1

03Identifiers

Citation
U-2 OS-ZFN-SNAP-Nup107(Cytion 目录号 300294)
NCBI TaxID
9606
Cellosaurus
CVCL_B7FM
Depositor
艾伦伯格实验室(EMBL)

04Expression and susceptibility

Protein expression
SNAP-Nup107(核孔复合体蛋白 107,SNAP-标记)

05Handling and culture

Culture medium
McCoys 5a,w:3.0 克/升葡萄糖,w:稳定谷氨酰胺,w:2.0 毫摩尔丙酮酸钠,w:2.2 克/升 NaHCO3(Cytion 文章编号 820200a)。
Incubation atmosphere

37°C, 5%CO2, 加湿环境。

Supplements
在培养基中添加 10% FBS、3.0 g/L 葡萄糖、稳定的谷氨酰胺、2.0 mM 丙酮酸钠、2.2 g/L NaHCO3、1% NEAA
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.

06Quality 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。

07Donor

Donor age
15 年
Ethnicity
高加索人