
- Tissue
- 胸腔积液
- Disease
- 多发性骨髓瘤
- Synonyms
- KMS 12 PE, KMS-12_PE, KMS-12PE, KMS12-PE, KMS12PE, Kawasaki Medical School-12-Pleural Effusion
Documents
KMS-12-PE
Cell line · Product number 300286
The KMS-12-PE cell line, established from the pleural effusion of the same patient, differs significantly from KMS-12-BM in several aspects. KMS-12-PE cel...
Recommended (freshest lot)
01General information
- Description
从同一患者胸腔积液中提取的 KMS-12-PE 细胞系在多个方面与 KMS-12-BM 有明显不同。KMS-12-PE细胞代表了分化更晚期的浆细胞阶段,CD20缺失,但CD38和PCA-1继续表达。KMS-12-PE 的一个显著特点是,无论是在患者胸腔积液中还是在培养过程中,它都能异位产生和分泌唾液型淀粉酶,这使它成为人类骨髓瘤细胞系中独一无二的细胞。这种现象与淀粉酶基因所在区域附近的染色体缺失有关,特别是在相当大比例的 KMS-12-PE 细胞中观察到的 del(1)(p22→pter)。
尽管存在这些明显的差异,KMS-12-PE 和 KMS-12-BM 都具有相同的克隆标记,即骨髓瘤病例中常见的 t(11;14)(q13;q32)易位。不过,KMS-12-PE 细胞的染色体异常少于 KMS-12-BM,而且往往是低二倍体。与KMS-12-BM一样,KMS-12-PE也不产生表面或分泌型免疫球蛋白,尽管细胞具有发达的内质网。尽管这两种细胞系在体外生长具有侵袭性,但它们都没有致瘤性,而且在无血清培养基中长期稳定增殖,这使它们成为研究骨髓瘤生物学的重要工具,尤其是在不产生免疫球蛋白的骨髓瘤中。
- Tissue
- 胸腔积液
- Disease
- 多发性骨髓瘤
- Synonyms
- KMS 12 PE
- KMS-12_PE
- KMS-12PE
- KMS12-PE
- KMS12PE
- Kawasaki Medical School-12-Pleural Effusion
02Classification
- Organism
- Human
- Sex
- Female
- Growth properties
- Suspension
- Biosafety level
- 1
03Characteristics
- Morphology
- 圆形电池
- Cell type
- B 细胞
- Tumourigenic
- 对裸鼠无致瘤性
04Identifiers
- Citation
- KMS-12-PE(Cytion 目录号 300286)
- NCBI TaxID
- 9606
- Cellosaurus
- CVCL_1333
05Expression and susceptibility
- Surface antigens
- cd3 -, cd4 -, cd13 -, cd14 -, cd15 -, cd19 -, cd20 -, cd34 -, cd38 +, cd138 +, hla-dr +, pca-1 +
- Mutational profile
- 易位:t(11;14)(q13;q32)
- Products
- 不产生免疫球蛋白
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
- Subculturing
- 通过定期添加或更换培养基来维持细胞培养。初始培养时细胞密度应为5×10⁵个细胞/毫升,并保持细胞浓度在3×10⁵至1×10⁶个细胞/毫升范围内以获得最佳生长效果。
- Seeding density
- 5 × 10⁵ 个细胞/毫升
- 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
- Donor age
- 64 岁
- Ethnicity
- 日语
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。
- 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