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B-LCL-CDG5
Tissue
外周血
Disease
EBV转化B淋巴母细胞样细胞系(B-LCL);PMM2-CDG(先天性糖基化障碍,源自患者)
Metastatic site
不适用(EBV转化性B细胞淋巴细胞样细胞瘤;无转移)

Documents

B-LCL-CDG5

Cell line · Product number 302016

B-LCL-CDG5 is an EBV-transformed B lymphocyte cell line derived from a patient with PMM2-CDG, a congenital disorder of glycosylation (CDG) caused by mutat...

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01General information

Description

B-LCL-CDG5是一种EBV转化的B淋巴细胞系,来源于一名PMM2-CDG患者,PMM2*基因突变导致先天性糖基化紊乱(CDG)。这种疾病会损害糖蛋白和糖脂上糖结构的正常合成和附着,影响多个器官系统。磷酸甘露糖突变酶2(PMM2)的缺乏会破坏甘露糖-6-磷酸向甘露糖-1-磷酸的转化,而甘露糖-1-磷酸是糖基化过程中的一个关键步骤,从而导致细胞功能缺陷和全身并发症。

B-LCL-CDG5作为一种EBV-immortalized B细胞系,是研究*PMM2*突变的生化和分子效应的重要模型。这种细胞系使研究人员能够研究糖基化缺陷、PMM2 酶活性以及糖基化受损的细胞后果。此外,它还为测试潜在的治疗方法(如药理伴侣、酶增强疗法或底物补充策略)提供了一个平台。B-LCL-CDG5与其他CDG患者衍生细胞系相结合,有助于增进我们对PMM2-CDG的了解,并开发有针对性的治疗方案。

Tissue
外周血
Disease
EBV转化B淋巴母细胞样细胞系(B-LCL);PMM2-CDG(先天性糖基化障碍,源自患者)
Metastatic site
不适用(EBV转化性B细胞淋巴细胞样细胞瘤;无转移)
Applications
免疫细胞中 CDG 作用的基因分型、功能测试(如 B 细胞表面抗原)、细胞毒性药物测试。突变分析、细胞凋亡机制分析、HLA 分型、不同细胞糖蛋白的糖基化缺陷对不同功能的影响。

02Classification

Organism
Human
Sex
Female
Growth properties
Suspension
GMO status
GMO-S2
GMO status details
GMO-S2: This B-LCL contains a stably maintained EBV episome encoding viral latent-phase genes (EBNA-1/-2/-3, LMP-1/-2). EBV is a risk group 2 pathogen; BSL-2 containment required. This classification applies within Germany; regulations may differ elsewhere.
Biosafety level
2

03Characteristics

Morphology
圆形电池
Cell type
B 淋巴细胞

04Identifiers

Citation
B-LCL-CDG5(Cytion 目录号 302016)
NCBI TaxID
9606
Cellosaurus
Not assigned

05Expression and susceptibility

Viruses
变形体:EBV

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% 热灭活 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2×10⁵个细胞/毫升,并保持细胞浓度在1×10⁵至5×10⁵个细胞/毫升范围内以获得最佳生长效果。
Fluid renewal
一旦中等颜色变成黄色
Post-thaw recovery
中型
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

Ethnicity
高加索人