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HEK293T/17
Tissue
胚胎肾脏
Synonyms
HEK293T/17, HEK-293T/17, HEK 293T/17

Documents

HEK293T/17

Cell line · Product number 305117

The 293T/17 cell line is an immortalized variant of the HEK293 line, derived from human embryonic kidney cells and extensively utilized in research, parti...

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01General information

Description

293T/17 细胞系是 HEK293 细胞系的永生化变种,源自人类胚胎肾细胞,广泛用于研究,特别是研究和生产逆转录病毒和慢病毒载体。该细胞系经改造后可表达 SV40 大 T 抗原,从而提高了其在病毒载体生产中的实用性。SV40 大 T 抗原的表达是这些细胞能够复制含有 SV40 复制源的质粒的一个关键特征,从而大大提高了瞬时转染程序中质粒 DNA 的产量。这一特性尤其有利于病毒载体的生产。

293T/17 细胞是生产逆转录病毒和慢病毒等病毒载体的关键。由于 293T/17 细胞能够扩增转染质粒并支持病毒的组装和释放,因此能高效地生产病毒颗粒。这使它们成为基因治疗研究的重要工具,这些载体被用于将遗传物质送入宿主细胞。细胞表现出很高的转染效率,这对于在载体构建过程中成功引入和表达外来 DNA 至关重要。这种高效率可以有效地研究基因功能和生成重组蛋白。

293T/17 细胞系的强大功能使其在基础科学研究和治疗应用中都具有重要价值。它广泛应用于分子生物学和基因工程中的蛋白质表达、基因功能分析和新型基因疗法的开发。该细胞系在生产病毒载体方面的高效率为需要遗传物质传递的实验提供了便利,使其成为病毒学领域的基石。293T/17 细胞系在促进我们对基因功能的了解和开发治疗干预措施方面继续发挥着关键作用。

Tissue
胚胎肾脏
Applications
这种细胞系是转染、高通量筛选、毒理学和疫苗开发的最佳选择。
Synonyms
  • HEK293T/17
  • HEK-293T/17
  • HEK 293T/17

02Classification

Organism
Human
Sex
Female
Growth properties
Adherent
GMO status
GMO-S1
GMO status details
GMO-S1: This HEK293T/17 cell line contains SV40, enhancing plasmid replication and packaging efficiency. The insert is stably present in transformed embryonic kidney cells. This classification applies only within Germany and may differ elsewhere.
Biosafety level
1

03Characteristics

Morphology
上皮细胞

04Identifiers

Citation
HEK293T/17(Cytion 产品编号 305117)
NCBI TaxID
9606
Cellosaurus
CVCL_1926

05Expression and susceptibility

Antigen expression
SV40 T 抗原
Viruses
SV40(表达 SV40 T 抗原)

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
胎儿