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MV3
Tissue
皮肤
Disease
黑色素瘤
Metastatic site
淋巴结
Synonyms
MV-3, MV 3

Documents

MV3

Cell line · Product number 305495

The MV3 cell line is a human melanoma model established from a metastatic lymph node of a male patient with amelanotic melanoma. This cell line is highly ...

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01General information

Description

MV3细胞系是一种从一名患有无色素性黑色素瘤的男性患者的转移性淋巴结中建立的人类黑色素瘤模型。该细胞系具有很强的转移性和致瘤性,因此成为研究黑色素瘤进展和转移的关键资源。 MV3 细胞表现出显著的侵袭性,这得益于其强大的降解细胞外基质成分的能力。这种能力通过尿激酶型纤溶酶原激活物(u-PA)通路介导,并受纤溶酶原激活物抑制剂(PAI-1 和 PAI-2)的调节。 MV3细胞的特征在于其表达与转移相关的标志物,例如整合素(如VLA-2)和生长因子受体,这些标志物与其侵袭性表型密切相关。

在异种移植模型中,MV3细胞表现出快速的肿瘤形成能力和高频率的自发转移,尤其是向肺部的转移。这些特征在旨在阐明黑色素瘤扩散和耐药性机制的临床前研究中起着关键作用。 裸鼠体内的MV3肿瘤保留了源自患者的原始肿瘤的组织学特征,包括高有丝分裂活性和有限的色素沉着。MV3细胞的促凝血特性进一步促使其转移行为,因为它们表现出组织因子活性,并能直接将凝血酶原激活为凝血酶,从而增强与肿瘤微环境的相互作用。

MV3还被用于研究细胞表面分子及其调控在黑色素瘤转移中的作用。例如,该细胞系表达显著水平的细胞间粘附分子-1(ICAM-1)和表皮生长因子受体(EGFR),这些分子与增强的转移潜能相关。 该模型在评估靶向治疗以及阐明黑色素瘤恶性程度的分子驱动因素方面,依然发挥着关键作用。

Tissue
皮肤
Disease
黑色素瘤
Metastatic site
淋巴结
Synonyms
  • MV-3
  • MV 3

02Classification

Organism
Human
Sex
Male
Growth properties
Adherent
Biosafety level
1

03Characteristics

Morphology
上皮样

04Identifiers

Citation
MV3(Cytion 产品编号 305495)
NCBI TaxID
9606
Cellosaurus
CVCL_W280

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
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。
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
76 年
Ethnicity
未说明