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B-LCL-CDG7
Tissue
外周血
Disease
先天性糖基化紊乱

Documents

B-LCL-CDG7

Cell line · Product number 302018

B-LCL-CDG7 is an EBV-transformed B lymphocyte cell line derived from a young boy with CDAII. CDAII is a rare genetic anaemia, affiliated to the class of CDG ...

US$800.00excl. tax and shipping
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Lot
Recommended (freshest lot)
1

01General information

Description
B-LCL-CDG7是一种EBV转化的B淋巴细胞系,来源于一名患有CDAII的小男孩。CDAII 是一种罕见的遗传性贫血,属于 CDG 糖基化紊乱。CDAII 患者的 COPII 成分 SEC23B 基因有缺陷,该基因参与细胞内蛋白质转运系统(特别是从 ER 的囊泡出芽)。患者为该基因的同基因突变。红细胞膜的带3糖蛋白糖基化不足,糖蛋白的多半乳糖胺基团糖基化异常,而糖磷脂糖基化异常,因此CDA II红细胞的带3具有截短的杂交型寡糖。这表明高尔基体糖基化酶 Beta-mannosidase II 或 Nacetylglucosaminyltransferase II 存在额外缺陷。
Tissue
外周血
Disease
先天性糖基化紊乱
Applications
免疫细胞 CDG 影响的基因分型、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、不同细胞糖蛋白糖基化缺陷对不同功能的影响。

02Classification

Organism
Human
Sex
Male
Growth properties
Suspension
Biosafety level
2

03Characteristics

Morphology
圆形电池
Cell type
B 淋巴细胞

04Identifiers

Citation
B-LCL-CDG7(Cytion 目录号 302018)
NCBI TaxID
9606
Cellosaurus
CVCL_A9Y3

05Expression and susceptibility

Surface antigens
CD15(Lewis x)(+)、CD15s(硅氨酰化 Lewis x)-、CD75s(硅氨酰化乳氨酰诺寡糖)+、CD173(血型 H)-、CD174(血型 Lewis y)-、CD175(Tn)-、CD175s(硅氨酰化 Tn)-、CD176(TF)+
Antigen expression
CD19+、CD20+、CD37+、CD43+、CD44+、CD45+、CD45R0-MHC I 类+、MHC II 类 (HLA-DR)+
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

Donor age
儿童
Ethnicity
高加索人