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- Tissue
- 造血
- Disease
- 骨髓瘤
- Synonyms
- P3-x63-Ag8.653, P3-x63-Ag8-653, P3-x63-Ag8 653, P3-x63-Ag 8.653, P3-x63Ag8.653, P3-x63.Ag8.653, P3/x63/Ag8.653, P3x63 Ag8.653, P3x63 AG8-653, P3x63-Ag8.653, P3x63-Ag8.653, P3x63 AG 8.653, P3x63Ag8653, P3-x63-Ag8-6-5-3, P3x63Ag8-6-5-3, P3.times.63 Ag8.653, P3.653, x63-Ag 8.6.5.3, x63-AG 8.653, x63-Ag8-653, x63-Ag8.653, x63.Ag8.653, x63Ag8-653, x63Ag8.653, x63AG8.653, P3-653, GM03570, GM3570, GM03570E, NS653
Documents
P3x63Ag8.653
Cell line · Product number 400118
The cells are resistant to 8-azaguanine and are HAT sensitive. They can be used as fusion partners for producing hybridomas. The cells do not secrete immunog...
Recommended (freshest lot)
01General information
- Description
- 这种细胞对 8-氮杂鸟嘌呤有抗性,对 HAT 敏感。它们可用作生产杂交瘤的融合伙伴。这种细胞不分泌免疫球蛋白。据报道,由于缺乏 3-酮固醇还原酶活性,这种细胞具有胆固醇辅助性。
- Tissue
- 造血
- Disease
- 骨髓瘤
- Synonyms
- P3-x63-Ag8.653
- P3-x63-Ag8-653
- P3-x63-Ag8 653
- P3-x63-Ag 8.653
- P3-x63Ag8.653
- P3-x63.Ag8.653
- P3/x63/Ag8.653
- P3x63 Ag8.653
- P3x63 AG8-653
- P3x63-Ag8.653
- P3x63-Ag8.653
- P3x63 AG 8.653
- P3x63Ag8653
- P3-x63-Ag8-6-5-3
- P3x63Ag8-6-5-3
- P3.times.63 Ag8.653
- P3.653
- x63-Ag 8.6.5.3
- x63-AG 8.653
- x63-Ag8-653
- x63-Ag8.653
- x63.Ag8.653
- x63Ag8-653
- x63Ag8.653
- x63AG8.653
- P3-653
- GM03570
- GM3570
- GM03570E
- NS653
02Classification
- Organism
- Mouse
- Sex
- Female
- Growth properties
- Adherent and suspension
- Biosafety level
- 1
03Characteristics
- Morphology
- 圆形电池
04Identifiers
- Citation
- P3x63Ag8.653(Cytion 目录编号 400118)
- NCBI TaxID
- 10090
- Cellosaurus
- CVCL_4032
05Expression and susceptibility
- Viruses
- 小鼠痘病毒检测呈阴性。
06Handling and culture
- Culture medium
- RPMI 1640,w:2.0 mM 稳定谷氨酰胺,w:2.0 g/L NaHCO3(Cytion 文章编号 820700a)。
- Incubation atmosphere
37°C, 5%CO2, 加湿环境。
- Supplements
- 在培养基中添加 10% FBS
- Dissociation reagent
- Accutase
- Subculturing
- 将悬浮细胞收集到 15 毫升试管中,用不含钙和镁的 PBS(T25 烧瓶用 3-5 毫升,T75 烧瓶用 5-10 毫升)轻轻清洗粘附的细胞。涂抹 Accutase(T25 烧瓶 1-2 毫升,T75 烧瓶 2.5 毫升),确保完全覆盖细胞层。让细胞在室温下孵育 10 分钟。孵育后,将悬浮细胞和粘附细胞混合并离心。离心后,小心重悬细胞团,并将细胞悬浮液转移到装有新鲜培养基的新烧瓶中。
- Seeding density
- 以4×10⁵细胞/毫升的浓度启动新培养。细胞密度不应超过2×10⁶细胞/毫升。
- Fluid renewal
- 每隔 3 到 4 天。收集漂浮的细胞,离心后与新鲜培养基一起加入烧瓶中。
- Post-thaw recovery
- 解冻后,将细胞以5×10⁴个细胞/平方厘米的密度接种于培养皿中,让细胞从冷冻过程中恢复并至少贴壁48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.
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
- Breed / subspecies
- BALB/c
09Legal 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。
- Disclaimer
- 我们提供的细胞仅用于体外实验室研究目的,不用于临床或诊断用途,也不得用于人类或兽医目的。用户必须遵守在研究环境中处理和使用这些细胞的所有适用准则和法规。
- 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