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KHM-5M
Tissue
甲状腺
Disease
甲状腺无细胞癌
Metastatic site
胸腔积液
Synonyms
KHM/5M, KHM5M

Documents

KHM-5M

Cell line · Product number 305148

The KHM-5M cell line is an important model derived from a patient with undifferentiated thyroid carcinoma complicated by neutrophilia and malignant pleuri...

US$800.00excl. tax and shipping
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Recommended (freshest lot)
  • CoA
1

01General information

Description

KHM-5M细胞系是一个重要的模型,它来源于一名并发中性粒细胞增多症和恶性胸膜炎的未分化甲状腺癌患者。这种细胞系的特点是能大量产生中性粒细胞趋化因子,特别是人白细胞介素8(IL-8)和粒细胞-巨噬细胞集落刺激因子(GM-CSF)。这些因子对中性粒细胞的招募和活化至关重要,而中性粒细胞在免疫反应和炎症中发挥着关键作用。研究表明,KHM-5M 细胞具有极强的趋化活性,体外实验中使用该细胞的条件培养基和改良的波登室技术证实了这一特性。

此外,将 KHM-5M 细胞移植到裸鼠体内,观察到中性粒细胞在移植的肿瘤组织内和周围浸润。这一发现强调了 KHM-5M 作为研究肿瘤细胞与免疫微环境之间相互作用的模型的相关性,特别是与中性粒细胞招募和功能有关的方面。该细胞系也是研究癌症细胞因子产生及随后改变病理特征的分子机制的重要工具。通过DNA克隆技术,证实了IL-8和GM-CSF的趋化活性,从而巩固了KHM-5M细胞系作为细胞因子驱动的肿瘤免疫相互作用研究的重要资源的地位。

Tissue
甲状腺
Disease
甲状腺无细胞癌
Metastatic site
胸腔积液
Synonyms
  • KHM/5M
  • KHM5M

02Classification

Organism
Human
Sex
Male
Growth properties
Adherent
Biosafety level
1

03Characteristics

Morphology
成纤维细胞
Doubling time
27 小时

04Identifiers

Citation
KHM-5M(Cytion 产品编号 305148)
NCBI TaxID
9606
Cellosaurus
CVCL_2975

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。
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
65 岁