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B-LCL-HROC32 (Bc HROC32)
Tissue
外周血
Disease
癌症
Synonyms
B-LCL CO32, Bc HROC32

Documents

B-LCL-HROC32 (Bc HROC32)

Cell line · Product number 300862

B-LCL-HROC32 (Bc HROC32) is an Epstein-Barr virus (EBV)-immortalized human B lymphoblastoid cell line established from B lymphocytes isolated from either ...

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01General information

Description

B-LCL-HROC32(Bc HROC32)是一种由成人患者肿瘤组织或外周血中分离的B淋巴细胞建立的、经爱泼斯坦-巴尔病毒(EBV)永生化的人B淋巴母细胞样细胞系。 该细胞系通过体外感染获得:在环孢素A抑制T细胞与NK细胞增殖的条件下,使用源自B95/8松鼠猴细胞系的含EBV上清液进行感染。经数周培养后,成功获得稳定的淋巴母细胞增殖,形成持续增殖的单克隆或寡克隆B细胞群体,适用于长期体外扩增。

免疫表型分析显示,B-LCL-HROC32(Bc HROC32)呈现成熟活化的B细胞特征,表达CD19和CD20,同时高表达CD23、CD80等活化与成熟标志物。强烈的MHC I类和II类分子表达表明其抗原呈递功能完好。 根据克隆个体差异,可观察到CD27、CD38或CD138等分化相关标记物的可变表达,反映B细胞成熟的不同阶段。该细胞对T细胞标记物呈阴性,证实其谱系特异性。

功能上,B-LCL-HROC32(Bc HROC32)可分泌特定亚型的免疫球蛋白(如IgG、IgM或IgA),且在长期培养中保持稳定。分泌的抗体可从培养上清液中收集,用于下游应用,包括抗原结合检测、肿瘤细胞识别研究或疾病相关抗原鉴定。 作为EBV永生化B细胞模型,B-LCL-HROC32(Bc HROC32)为研究体液免疫应答、B细胞活化与分化、以及肿瘤免疫学或系统性免疫应答中的抗体介导机制提供了强大的体外研究平台。

Tissue
外周血
Disease
癌症
Synonyms
  • B-LCL CO32
  • Bc HROC32

02Classification

Organism
Human
Sex
Female
Growth properties
Suspension
Biosafety level
2

03Characteristics

Morphology
圆形电池
Cell type
B 淋巴细胞

04Identifiers

Citation
B-LCL-HROC32 (Bc HROC32)(Cytion 目录号 300862)
NCBI TaxID
9606
Cellosaurus
CVCL_YD55

05Expression and susceptibility

Surface antigens
CD19
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
通过上下吸取操作轻柔匀化烧瓶中的细胞悬液,随后取代表性样本测定每毫升细胞密度。用新鲜培养基将悬液稀释至1×10⁵细胞/毫升浓度,并将调整后的悬液分装至新烧瓶中进行后续培养。
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
82 年
Ethnicity
高加索人