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HARA-B
Tissue
肺部
Disease
肺鳞癌
Metastatic site
胸腔积液
Synonyms
HARAB

Documents

HARA-B

Cell line · Product number 300465

The HARA-B cell line is derived from human squamous cell carcinoma of the lung, specifically established from metastatic bone tissue in a mouse model. Thi...

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01General information

Description

HARA-B 细胞系来源于人类肺鳞状细胞癌,特别是从小鼠模型的转移性骨组织中建立的。该细胞系是由最初的 HARA 细胞系二次开发而成,其特点是高表达甲状旁腺激素相关蛋白 (PTHrP),PTHrP 在这些细胞中观察到的广泛骨转移中起着重要作用。HARA-B 细胞系在研究与肺癌相关的骨转移机制方面发挥了重要作用。

涉及 HARA-B 的科学研究通常关注其在高钙血症模型中的作用,高钙血症是与包括肺癌在内的某些癌症相关的常见副肿瘤综合征。该模型中的高钙血症是通过皮下注射移植细胞诱发的,为了解癌细胞与骨细胞之间的相互作用以及导致骨降解和钙释放的途径提供了宝贵的工具。这种细胞系有助于研究人员研究潜在的治疗策略,以减轻肺癌患者的骨转移和相关并发症。

Tissue
肺部
Disease
肺鳞癌
Metastatic site
胸腔积液
Synonyms
  • HARAB

02Classification

Organism
Human
Sex
Male
Growth properties
Adherent

03Identifiers

Citation
HARA-B(Cytion 目录编号 300465)
NCBI TaxID
9606
Cellosaurus
CVCL_2915

04Expression and susceptibility

Protein expression
产生大量甲状旁腺激素相关肽(PTHrP)。

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
57 年
Ethnicity
日语