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WIL2
Tissue
脾
Disease
遗传性球形红细胞症
Synonyms
WIL-2, Wil.2, WI-L2, Wi-L2

Documents

WIL2

Cell line · Product number 302011

Wil2 is a human B lymphoblastoid cell line derived from peripheral blood B lymphocytes of an adult donor and subsequently immortalized through Epstein–Bar...

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01General information

Description

Wil2 是一种源自成年供体外周血 B 淋巴细胞的人 B 淋巴母细胞样细胞系,随后通过爱泼斯坦-巴尔病毒(EBV)转化而获得永生化。 作为一种EBV阳性的悬浮细胞系,Wil2表现出活化B细胞的特征性表现,包括持续增殖、表达B细胞表面标志物以及合成免疫球蛋白的能力。这些细胞以单细胞或小簇形式在悬浮液中生长,通常在添加血清的标准淋巴细胞培养条件下维持。

表型上,Wil2细胞表达典型的B细胞谱系标志物(如CD19、CD20和表面免疫球蛋白),同时还表达由EBV潜伏基因表达诱导的激活相关标志物。 EBV环状DNA的存在驱动细胞增殖并支持长期培养,使该细胞系成为研究病毒潜伏、B细胞激活及宿主-病毒相互作用的有用模型。此外,Wil2已被用于免疫学和分子生物学研究,重点关注转化B淋巴细胞中的抗体产生、抗原呈递及信号转导通路。

尽管Wil2是典型的EBV转化B细胞模型,但与其相比,针对其详细遗传背景和功能特异性的已发表数据仍相对有限,而其他淋巴母细胞样细胞系的表征则更为详尽。建议研究人员在各自的实验背景下验证具体的表型或功能特性,并查阅最新数据库或原始文献以获取最前沿的表征数据。

Tissue
脾
Disease
遗传性球形红细胞症
Synonyms
  • WIL-2
  • Wil.2
  • WI-L2
  • Wi-L2

02Classification

Organism
Human
Sex
Male
Growth properties
Suspension
Biosafety level
1

03Characteristics

Cell type
B 淋巴细胞
Karyotype
46、低二倍体

04Identifiers

Citation
WIL2(Cytion 目录号 302011)
NCBI TaxID
9606
Cellosaurus
CVCL_6544

05Handling 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% 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5×10⁵个细胞/毫升,并保持细胞浓度在3×10⁵至1×10⁶个细胞/毫升范围内以获得最佳生长效果。
Seeding density
1 × 10⁵ 个细胞/毫升
Fluid renewal
每周 2 次
Post-thaw recovery
快速
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.

06Quality 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。

07Donor

Donor age
5 年
Ethnicity
高加索人