HSF(SV40)
Cell line · Product number 305338
The HSF(SV40) immortalized cell line refers to cells that have been genetically modified to express the Simian Virus 40 (SV40) large T-antigen (T-Ag), whi...
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01General information
- Description
HSF(SV40)永生化细胞系是指经过基因改造的细胞,这些细胞表达能促进细胞永生化的 Simian Virus 40 (SV40) 大 T 抗原 (T-Ag)。SV40 T-Ag 是一种强效的肿瘤蛋白,能与 p53 和视网膜母细胞瘤蛋白(Rb)等重要的肿瘤抑制蛋白相互作用,导致其抑制肿瘤的功能失活。这种相互作用破坏了正常的细胞周期控制机制,使细胞得以绕过衰老,无限增殖。
由于 HSF(SV40)细胞具有永生的特性,而且 SV40 T-Ag 在其转化过程中起着关键作用,因此被广泛用于癌症研究,特别是与病毒致癌、细胞周期调控以及针对分子伴侣和肿瘤抑制途径的治疗干预相关的研究。它们的使用为了解病毒癌蛋白与宿主细胞调控网络之间的相互作用提供了宝贵的见解,为开发癌症靶向疗法铺平了道路。
- Metastatic site
- 不适用(正常脾脏成纤维细胞;SV40永生化)
- Applications
- SV40致癌作用;p53/Rb肿瘤抑制基因生物学;细胞周期调控;成纤维细胞生物学;癌症生物学
02Classification
- Organism
- Human
- Growth properties
- Adherent
- GMO status
- GMO-S1
- GMO status details
- GMO-S1: This HSF fibroblast line contains an SV40 T-antigen construct enabling immortalization for dermal and connective tissue studies. This classification applies only within Germany and may differ elsewhere.
- Biosafety level
- 1
03Characteristics
- Morphology
- 纤维母细胞样
- Cell type
- 成纤维细胞
- Doubling time
- 约24至36小时
04Identifiers
- Citation
- HSF(SV40)(Cytion 目录号 305338)
- NCBI TaxID
- 9606
- Cellosaurus
- Not assigned
05Handling 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
- 用PBS洗涤,加入Accutase(室温下孵育8–10分钟),重新悬浮,以300×g离心3分钟,然后重新接种。
- Split ratio
- 1 至 3
- Seeding density
- 1 至 3 × 10⁴ 个细胞/cm²
- 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 300 x g for 3 minutes to separate the cells and carefully discard the supernatant containing residual freezing medium.
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.
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。
07Legal 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
