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HEL
Tissue
外周血
Disease
红细胞白血病
Synonyms
Hel, GM06141, GM06141B, Human ErythroLeukemia

Documents

HEL

Cell line · Product number 305022

HEL cells are a human erythroleukemia cell line that was established from the peripheral blood of a 30-year-old man with erythroleukemia in relapse after ...

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01General information

Description

HEL 细胞是一种人类红白血病细胞系,是 1980 年从一名 30 岁的红白血病患者的外周血中提取的。

HEL 细胞能够自发合成和诱导合成球蛋白,主要产生 G γ 和 A γ 链。这些细胞还表达极少量的胚胎链(epsilon、zeta)和α链,而β链则检测不到。

HEL 细胞呈圆形,大至偶见巨型多核悬浮单细胞,少数细胞粘附。通过 RT-PCR 和测序,已证实这些细胞中表达了突变的 JAK2。HEL 细胞表达多种细胞表面标志物,包括 CD3-、CD13+、CD14-、CD19-、CD33+、CD41a+、CD71+ 和 CD235a+。 根据研究,羟基脲是治疗包括红细胞白血病在内的多种癌症的常规药物,它也可能调节 HEL 细胞的死亡。

羟基脲导致的 HEL 细胞凋亡可能与 HEL 细胞的末端分化有关。此外,早前的研究表明,羟基脲可能在控制 HEL 细胞增殖和分化方面起着关键作用。

Tissue
外周血
Disease
红细胞白血病
Synonyms
  • Hel
  • GM06141
  • GM06141B
  • Human ErythroLeukemia

02Classification

Organism
Human
Sex
Male
Growth properties
Suspension
Biosafety level
1

03Characteristics

Morphology
圆形
Doubling time
36 小时

04Identifiers

Citation
HEL(Cytion 产品编号 305022)
NCBI TaxID
9606
Cellosaurus
CVCL_0001

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
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。
Fluid renewal
每周 2 至 3 次
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
30年
Ethnicity
欧洲