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HuH7 细胞
Tissue
肝脏
Disease
肝细胞癌
Metastatic site
肝癌
Synonyms
HuH-7, HUH-7, Huh-7, Huh7, HUH7, HUH7.0, JTC-39, Japanese Tissue Culture-39

Documents

HuH7 细胞

Cell line · Product number 300156

HuH-7 cells are a type of epithelial-like, tumorigenic cell line initially taken from a liver tumor in a 57-year-old Japanese male in 1982. The human hepa...

US$550.00excl. tax and shipping
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Lot
Recommended (freshest lot)
  • CoA
1

01General information

Description

HuH-7细胞是一种上皮样肿瘤细胞系,最初于1982年从一名57岁的日本男性肝脏肿瘤中提取。人肝癌衍生的 HuH-7 细胞系及其衍生物作为原代肝细胞的方便实验替代品已被广泛用于研究。特别是,它们在丙型肝炎研究中发挥了重要作用,并被用作体外繁殖病毒的宿主细胞。HuH-7 细胞在丙型肝炎研究中发挥了至关重要的作用,尤其是在药物开发方面。2005 年以前,研究人员无法在实验室中培养丙型肝炎病毒,因此很难测试针对丙型肝炎病毒的潜在候选药物。

HuH-7 细胞系的问世改变了这一状况。这些细胞对丙型肝炎病毒的复制具有高度的容许性,因此非常适合进行体外测试。通过使用 HuH-7 细胞,研究人员能够筛选出针对实验室培养的丙型肝炎的候选药物,这为开发抗病毒新药铺平了道路。与其他已建立的人类肝癌细胞系不同,HuH-7 细胞可以在含有微量硒的化学定义培养基中代替血清进行繁殖。这样就可以系统地研究各种化合物对其生长和新陈代谢的体外影响。

Tissue
肝脏
Disease
肝细胞癌
Metastatic site
肝癌
Synonyms
  • HuH-7
  • HUH-7
  • Huh-7
  • Huh7
  • HUH7
  • HUH7.0
  • JTC-39
  • Japanese Tissue Culture-39

02Classification

Organism
Human
Sex
Male
Growth properties
Adherent
Biosafety level
1

03Characteristics

Morphology
上皮样
Tumourigenic
是的,在裸鼠身上。
Doubling time
48 小时

04Identifiers

Citation
HuH7(Cytion 目录号 300156)
NCBI TaxID
9606
Cellosaurus
CVCL_0336

05Expression and susceptibility

Viruses
人乳头瘤病毒(HPV)、丙型肝炎病毒(HCV)和艾滋病毒(HIV)阴性。

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% 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
常规细胞培养期间为1至2×10⁴个细胞/平方厘米
Fluid renewal
每 3 天
Post-thaw recovery
以2至3×10⁴个细胞/平方厘米的密度开始培养。细胞将在24至48小时内恢复。
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
57 年
Ethnicity
日语