CLS-439 细胞
Cell line · Product number 300150
Established from the primary bladder carcinoma of a 61-year-old male in 1998 by CLS.
Recommended (freshest lot)
01General information
- Description
- 1998 年通过 CLS 从一名 61 岁男性的原发性膀胱癌中确诊。
- Tissue
- 膀胱
- Disease
- 癌症
- Synonyms
- CLS439
02Classification
- Organism
- Human
- Sex
- Male
- Growth properties
- Adherent
- Biosafety level
- 1
03Characteristics
- Morphology
- 上皮样
- Tumourigenic
- 是,在裸鼠身上
- Doubling time
- 35 小时
04Identifiers
- Citation
- CLS-439(Cytion 目录号 300150)
- NCBI TaxID
- 9606
- Cellosaurus
- CVCL_5982
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
- Dissociation reagent
- Accutase
- Subculturing
- 去除培养基,用不含钙和镁的 PBS 冲洗粘附的细胞(T25 细胞培养皿用 3-5 毫升 PBS,T75 细胞培养皿用 5-10 毫升)。加入 TrypleExpress(每个 T25 细胞培养瓶 1-2 毫升,每个 T75 细胞培养瓶 2.5 毫升),细胞片必须完全覆盖。在环境温度下孵育 10 分钟。小心重悬细胞,可选择添加培养基,但不是必须添加,然后分装到装有新鲜培养基的新培养瓶中。
- Seeding density
- 1×10⁴个细胞/cm²将在约3天内形成融合细胞层
- Fluid renewal
- 每周 2 至 3 次
- Post-thaw recovery
- 细胞解冻后必须在 37 摄氏度/5%CO2的环境中静置至少 24 小时。
- 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。
07Donor
- Donor age
- 61 年
- Ethnicity
- 高加索人
08Legal 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
