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HEp-2
Tissue
喉部
Disease
腺癌
Synonyms
Hep-2, HEP-2, HEp-2/HeLa, Hep 2, Hep2, HEp2, HEP2, H.Ep.-2, H.Ep. #2, H.Ep. No. 2, Hep II, Human Epidermoid carcinoma #2, Human Epithelioma-2

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HEp-2

Cell line · Product number 300397

The HEp-2 cell line, originally believed to be derived from laryngeal cancer cells, was later identified through DNA fingerprinting and the presence of He...

US$395.00excl. tax and shipping
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Lot
Recommended (freshest lot)
  • CoA
1

01General information

Description

HEp-2 细胞系最初被认为来源于喉癌细胞,后来通过 DNA 指纹和 HeLa 标记染色体的存在被确认为受到 HeLa 细胞(一种来源于宫颈癌的细胞系)的污染。

尽管如此,HEp-2 细胞系仍被广泛用于间接免疫荧光法检测抗核抗体(ANA),这是诊断系统性红斑狼疮和系统性硬化症等疾病的关键。使用 HEp-2 细胞的间接免疫荧光检测法(IIFA)可提供明确的阳性或阴性结果,是检测抗核抗体的标准方法。这种直接的方法对于诊断和分类不同的系统性自身免疫疾病至关重要。

在 HEp-2 细胞间接免疫荧光法中观察到的自身抗体模式,尤其是在风湿病学中观察到的自身抗体模式,为了解各种风湿病提供了宝贵的信息。此外,对 HEp-2 人体细胞在不同培养条件下表达的抗原进行全面审查,有助于确定与狼疮等疾病相关的特异性 ANA。

总之,尽管 HEp-2 等细胞系受到 HeLa 细胞的污染,引发了癌症研究中对结果准确性和可靠性及其临床意义的担忧,但 Hep-2 在检测抗核抗体方面的实用性及其在不同研究学科中的应用凸显了其持续的重要性。除其他应用外,HEp-2 细胞系还是诊断和分类自身免疫性疾病的重要工具。

Tissue
喉部
Disease
腺癌
Applications
在风湿病学中,使用 HEp-2 细胞的间接免疫荧光在诊断自身免疫性疾病(包括系统性红斑狼疮和系统性硬化症)方面发挥着重要作用
Synonyms
  • Hep-2
  • HEP-2
  • HEp-2/HeLa
  • Hep 2
  • Hep2
  • HEp2
  • HEP2
  • H.Ep.-2
  • H.Ep. #2
  • H.Ep. No. 2
  • Hep II
  • Human Epidermoid carcinoma #2
  • Human Epithelioma-2

02Classification

Organism
Human
Sex
Female
Growth properties
Adherent
Biosafety level
1

03Characteristics

Morphology
上皮样

04Identifiers

Citation
HEp-2 (Cytion 目录号 300397)
NCBI TaxID
9606
Cellosaurus
CVCL_1906

05Expression and susceptibility

Isoenzymes
G6PD, A
Reverse transcriptase
阴性
Products
角蛋白

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 和 1% NEAA
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 2 至 3 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,使细胞从冷冻过程中恢复并至少贴壁24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
30年
Ethnicity
非洲裔美国人