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BC-3
Tissue
胸腔积液
Disease
原发性渗出性淋巴瘤
Synonyms
BC3

Documents

BC-3

Cell line · Product number 305748

The BC-3 cell line is a human primary effusion lymphoma (PEL) model derived from the pleural effusion of an HIV-negative patient. It represents a rare sub...

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01General information

Description

BC-3细胞系是一种源自HIV阴性患者胸腔积液的人原发性渗出性淋巴瘤(PEL)模型。它代表了一种罕见的B细胞非霍奇金淋巴瘤亚型,通常表现为胸膜、心包或腹膜等体腔内的淋巴瘤性积液,且往往未检测到实体肿瘤。 BC-3 的独特之处在于其仅感染卡波西肉瘤相关疱疹病毒(KSHV 或 HHV-8),且完全不携带埃博拉病毒(EBV),这一特征使其区别于大多数其他 PEL 细胞系——后者通常同时感染这两种病毒。

在免疫表型方面,BC-3细胞表达白细胞共同抗原(CD45),但不表达经典的B细胞(如CD19、CD20、CD21、CD22)和T细胞(如CD2、CD3、CD4、CD5、CD8)谱系标志物,这与PEL的未分化免疫表型一致。 然而,它们确实表现出 CD30、CD38、CD54 和 HLA-DR 等活化标志物的可变表达。在基因型方面,BC-3 表现出克隆性免疫球蛋白基因重排,表明其源自 B 细胞,但缺乏 c-myc 致癌基因重排。 重要的是,Southern印迹和PCR分析证实存在完整的约170 kb KSHV基因组,电子显微镜观察显示出疱疹病毒样衣壳,证实该细胞系是具有传染性的KSHV颗粒的有效来源。

BC-3已被纳入LL-100白血病和淋巴瘤细胞系库,并在转录组分析中与其他PEL模型形成独特的聚类。该细胞系表达PRDM1/BLIMP1、IL-10、SLAMF7以及S100A家族成员等PEL特征性基因。 这些标志物反映了生发中心后B细胞的表型,并突显了BC-3作为生物学相关模型在研究KSHV致病机制、PEL生物学特性以及针对KSHV相关恶性肿瘤的治疗策略方面的实用价值。

Tissue
胸腔积液
Disease
原发性渗出性淋巴瘤
Applications
免疫学
Synonyms
  • BC3

02Classification

Organism
Human
Sex
Male
Growth properties
Suspension
Biosafety level
2

03Characteristics

Morphology
淋巴母细胞
Doubling time
约50-60小时

04Identifiers

Citation
BC-3(Cytion 产品编号 305748)
NCBI TaxID
9606
Cellosaurus
CVCL_1080

05Expression and susceptibility

Antigen expression
CD30+;CD38+;CD45+;CD54+;CD71+;HLA-DR+;EMA+(上皮膜抗原);CD2-;CD3-;CD4-;CD5-;CD8-;CD19-;CD20-;CD21-;CD22-
Mutational profile
突变:基因缺失,CDKN2A,纯合子

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 20% 的 FBS
Dissociation reagent
无
Seeding density
2.5 至 10 × 10⁵ 个细胞/cm²
Fluid renewal
每周 2 至 3 次
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
作为冷冻保存培养基,我们使用完全生长培养基(含FBS)+ 5% DMSO以确保解冻后的细胞存活率,或使用CM-1(Cytion货号800100),该培养基含有经过优化的渗透保护剂和代谢稳定剂,可提高细胞恢复率并减轻冷冻引起的应激。
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
85 岁
Ethnicity
高加索人