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WI 38 VA13 subline 2RA
Tissue
肺部
Synonyms
WI 38 VA-13 subline 2RA, WI 38VA13 subline 2RA, WI-38 VA13 sub 2 RA, WI38-VA13 subline 2RA, WI38 VA13/2RA, WI38VA13/2RA, VA13 2RA, WI-38 VA13, WI 38 VA 13, WI38-VA13, WI38/VA13, WI38VA13, VA-13, VA13, AG07217, AG7217

Documents

WI 38 VA13 subline 2RA

Cell line · Product number 300421

The WI-38 VA13 subline 2RA, derived from the historic WI-38 cell line originally sourced from the lung tissue of a 3-month-old fetus, represents a key adv...

US$395.00excl. tax and shipping
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01General information

Description

WI-38 VA13 亚系 2RA 源自历史悠久的 WI-38 细胞系,该细胞系最初取自 3 个月大胎儿的肺组织,是细胞培养技术的一大进步。最初的 WI-38 细胞系在开发麻疹、流行性腮腺炎、风疹和甲型肝炎等多种病毒性疾病疫苗的过程中发挥了至关重要的作用。VA13 亚系 2RA 是该细胞系的永生化变种,是通过用 Simian Virus 40(SV40)转化实现的。

将 SV40 加入 WI-38 细胞以创建 VA13 亚系 2RA 延长了细胞的寿命,为长期实验提供了更持久的模型。这种转化保持了原始二倍体细胞的基本特性,但改变了它们的生命周期和生长模式,从而实现了持续生长,促进了广泛的研究,而这是母本细胞系有限的生命周期所无法实现的。这使得 VA13 亚系在病毒学、药理学和基因研究等需要延长观察期的持续和广泛研究领域特别有用。

Tissue
肺部
Synonyms
  • WI 38 VA-13 subline 2RA
  • WI 38VA13 subline 2RA
  • WI-38 VA13 sub 2 RA
  • WI38-VA13 subline 2RA
  • WI38 VA13/2RA
  • WI38VA13/2RA
  • VA13 2RA
  • WI-38 VA13
  • WI 38 VA 13
  • WI38-VA13
  • WI38/VA13
  • WI38VA13
  • VA-13
  • VA13
  • AG07217
  • AG7217

02Classification

Organism
Human
Sex
Female
Growth properties
Adherent
Biosafety level
2

03Characteristics

Morphology
上皮样
Cell type
成纤维细胞
Karyotype
超二倍体,模数:73-78

04Identifiers

Citation
WI 38 VA13 亚系 2RA(Cytion 目录号 300421)
NCBI TaxID
9606
Cellosaurus
CVCL_2759

05Expression and susceptibility

Isoenzymes
G6PD、B
Viruses
含巴氏病毒
Virus susceptibility
单纯疱疹、水泡性口炎(印第安纳)、脊髓灰质炎病毒 2
Reverse transcriptase
阴性

06Handling and culture

Culture medium
EMEM(MEM Eagle),w:2 mM L-谷氨酰胺,w:2.2 g/L NaHCO3,w:EBSS(Cytion 文章编号 820100a)。
Incubation atmosphere

37°C, 5%CO2, 加湿环境。

Supplements
在培养基中添加 10% FBS
Dissociation reagent
Accutase
Subculturing
去除粘附细胞上的旧培养基,用不含钙和镁的 PBS 冲洗。T25 烧瓶用 3-5 毫升 PBS,T75 烧瓶用 5-10 毫升。然后用 Accutase 完全覆盖细胞,T25 烧瓶用 1-2 毫升,T75 烧瓶用 2.5 毫升。让细胞在室温下孵育 8-10 分钟,使其分离。孵育后,用 10 毫升培养基轻轻混合细胞使其重悬,然后用 300xg 离心 3 分钟。弃去上清液,用新鲜培养基重悬细胞,并将其转移到已装有新鲜培养基的新烧瓶中。
Seeding density
1 × 10⁴ 个细胞/平方厘米
Fluid renewal
每周 1 至 2 次
Post-thaw recovery
解冻后,将细胞以5×10⁴个细胞/平方厘米的密度接种于培养皿中,让细胞从冷冻过程中恢复并至少贴壁48小时。
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
妊娠 3 个月
Ethnicity
高加索人