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MEG-01
Tissue
骨髓
Disease
慢性髓性白血病
Synonyms
Meg-01, MEG01, Meg01

Documents

MEG-01

Cell line · Product number 300482

The MEG-01 cell line is a human megakaryoblast cell line established from the bone marrow of a 55-year-old male patient who was in the megakaryoblastic cr...

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

01General information

Description

MEG-01 细胞系是从一名 55 岁男性慢性粒细胞白血病(CML)巨核细胞危象期患者的骨髓中建立的人类巨核细胞细胞系。该细胞系于 1983 年在日本名古屋大学医学院研制成功。提取 MEG-01 细胞的患者费城染色体(Ph1)呈阳性,这是慢性骨髓性白血病的标志。MEG-01 细胞表现出染色体数目模态为 56 至 58 的超二倍体核型,持续显示 Ph1 染色体的存在,这是 t(9;22)染色体易位的结果。

MEG-01 细胞具有混合生长特性,在培养过程中表现出粘附和悬浮两种特征。这些细胞表达巨核细胞系特有的几种标记和抗原,包括 CD41、CD61 和 CDw14。它们的细胞质因子 VIII、表面 GPIIb/IIIa 和各种酶活性(如周期性酸-希夫(PAS)反应、α-萘基乙酸酯酶和酸性磷酸酶)也呈阳性。有趣的是,MEG-01 细胞的髓过氧化物酶、α-萘基丁酸酯酶、萘酚 AS-D 氯代乙酸酯酶和碱性磷酸酶均为阴性,这有助于将它们与其他髓系细胞区分开来。

MEG-01 是研究人类巨核细胞生成、血小板生成和巨核细胞系特有蛋白质(如血小板衍生生长因子 (PDGF) 和 GPIIb/IIIa 等糖蛋白)生物合成的宝贵模型。由于 MEG-01 的遗传背景特征良好,而且能够表达关键的巨核细胞标志物,因此它是研究白血病和血小板生物生成机制的重要工具,尽管它并不打算用于治疗或体内应用。

Tissue
骨髓
Disease
慢性髓性白血病
Synonyms
  • Meg-01
  • MEG01
  • Meg01

02Classification

Organism
Human
Sex
Male
Growth properties
Adherent and suspension
Biosafety level
1

03Characteristics

Morphology
肌母细胞样
Cell type
巨核细胞

04Identifiers

Citation
MEG-01(Cytion 目录号 300482)
NCBI TaxID
9606
Cellosaurus
CVCL_0425

05Expression and susceptibility

Antigen expression
CD41 +、CD61 +、CDw14 +

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
将悬浮细胞收集到 15 毫升试管中,用不含钙和镁的 PBS(T25 烧瓶用 3-5 毫升,T75 烧瓶用 5-10 毫升)轻轻清洗粘附的细胞。涂抹 Accutase(T25 烧瓶 1-2 毫升,T75 烧瓶 2.5 毫升),确保完全覆盖细胞层。让细胞在室温下孵育 10 分钟。孵育后,将悬浮细胞和粘附细胞混合并离心。离心后,小心重悬细胞团,并将细胞悬浮液转移到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.

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
55 岁
Ethnicity
东亚