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B-LCL-HROG02
Tissue
外周血
Disease
EBV转化B淋巴母细胞样细胞系(B-LCL);胃癌相关供体

Documents

B-LCL-HROG02

Cell line · Product number 302106

B-LCL-HROG02 is a human Epstein-Barr virus (EBV)-transformed B lymphoblastoid cell line (B-LCL) established from the peripheral blood of a 68-year-old Cau...

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1

01General information

Description

B-LCL-HROG02 是一株由人类爱泼斯坦-巴尔病毒(EBV)转化而来的 B 淋巴母细胞样细胞系 (B-LCL),源自HROG生物样本库系列中一名68岁高加索裔男性胃癌患者的外周血,该系列是HROC结直肠癌B-LCL样本库的胃癌对应样本库。 通过EBV介导的原代外周血B淋巴细胞体外永生化,获得了一种具有淋巴母细胞样形态的稳定增殖悬浮细胞系。HROG02细胞持续表达EBV潜伏期抗原(EBNA-1、-2、-3、 LMP-1、-2),并高水平表达HLA I类和II类分子、共刺激分子CD80和CD86、黏附分子CD54和CD58,以及B细胞标志物CD19、CD20和CD23。

B-LCL-HROG02 适用于 T 细胞和 NK 细胞功能检测、HLA 分型、混合淋巴细胞反应、抗原呈递研究以及 CTL 诱导方案。 在HROG患者配对生物样本库中,该细胞系为胃癌研究提供了明确的免疫学参考标准,可用于开展肿瘤相关免疫应答的自体或异体研究。该细胞系支持对胃癌特异性T细胞反应性的研究,以及与胃肿瘤免疫学相关的患者配对免疫检测系统的研究。

Tissue
外周血
Disease
EBV转化B淋巴母细胞样细胞系(B-LCL);胃癌相关供体
Applications
免疫学研究;T细胞和NK细胞检测;HLA分型;抗原呈递研究;混合淋巴细胞反应;CTL检测靶细胞;胃癌免疫学

02Classification

Organism
Human
Sex
Male
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 classified as a risk group 2 pathogen. Handling requires BSL-2 containment. This classification applies within Germany; regulations may differ in other jurisdictions.
Biosafety level
2

03Characteristics

Morphology
淋巴母细胞样
Cell type
B 淋巴细胞
Doubling time
24 至 48 小时

04Identifiers

Citation
B-LCL-HROG02(Cytion 产品编号 302106)
NCBI TaxID
9606

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
在培养基中加入15–20%的FBS
Subculturing
每2–3天用新鲜预热的完全培养基将培养物稀释至2–5 × 10⁵个细胞/毫升。传代前计数活细胞,以维持最佳密度。 培养物浓度不得超过2 × 10⁶个细胞/毫升。必要时可通过离心(300 × g,5分钟)进行培养基更换。无需进行酶解分离。
Split ratio
1 至 4
Seeding density
2 至 5 × 10⁵ 个细胞/毫升
Fluid renewal
每 2 至 3 天
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 200 x g for 5 minutes, carefully discard the supernatant containing freezing medium.

  7. Follow the procedure described under Post-Thaw Recovery

07Quality control and cryopreservation

Freeze medium
作为冷冻保存培养基,我们使用完全生长培养基+10% DMSO,以获得足够的解冻后存活率。
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
68岁
Ethnicity
高加索人