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KHOS-240S
Tissue
骨质
Disease
骨肉瘤
Synonyms
KHOS240S

Documents

KHOS-240S

Cell line · Product number 300433

KHOS-240S is an osteosarcoma cell line that is derived from human bone sarcoma tissue. This cell line, along with its variants, has been extensively used ...

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01General information

Description

KHOS-240S 是一种骨肉瘤细胞系,来源于人类骨肉瘤组织。这种细胞系及其变体已被广泛用于骨肉瘤的研究,骨肉瘤是一种主要影响儿童和年轻人的原发性恶性骨肿瘤。骨肉瘤的特点是恶性细胞产生未成熟骨(类骨),因其侵袭性和早期转移的潜力而臭名昭著,尤其是转移到肺部。

KHOS-240S 细胞系对多种激酶抑制剂(包括针对 PI3K-Akt-mTOR 通路的抑制剂)具有抗药性。这种对常见治疗靶点的耐药性使得KHOS-240S对研究骨肉瘤的耐药机制和探索替代治疗策略特别有价值。研究人员利用这种细胞系筛选了多种肿瘤药物和研究药物,从而发现了有可能克服耐药机制的化合物。在利用 KHOS-240S 进行的研究中,与耐药性和骨肉瘤生物学相关的基因(如参与 mTOR 信号通路的基因)的表达谱尤其引人关注。

此外,KHOS-240S 还被用于探索 microRNA 的表达模式,这可能与药物敏感性或耐药性有关。该细胞系对 PI3K-Akt-mTOR 通路抑制剂的特异性耐药性为了解骨肉瘤如何逃避靶向治疗提供了一个重要模型,并为开发新型治疗方法提供了基础,从而提高耐药骨肉瘤亚型的治疗效果。

Tissue
骨质
Disease
骨肉瘤
Synonyms
  • KHOS240S

02Classification

Organism
Human
Sex
Female
Growth properties
Adherent
Biosafety level
1

03Characteristics

Morphology
纤维母细胞样
Tumourigenic
没有

04Identifiers

Citation
KHOS-240S(Cytion 目录号 300433)
NCBI TaxID
9606
Cellosaurus
CVCL_2544

05Handling and culture

Culture medium
EMEM(MEM Eagle),w:2 mM L-谷氨酰胺,w:2.2 g/L NaHCO3,w:EBSS(Cytion 文章编号 820100a)。
Incubation atmosphere

37°C, 5%CO2, 加湿环境。

Supplements
在培养基中添加 10% FBS 和 1% NEAA
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
1 × 10⁴ 个细胞/平方厘米
Fluid renewal
每周 2 至 3 次
Post-thaw recovery
解冻后,将细胞以5×10⁴个细胞/平方厘米的密度接种于培养皿中,使细胞从冷冻过程中恢复并至少贴壁24小时。
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
13 年
Ethnicity
高加索人