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KMH-2
Tissue
甲状腺
Disease
甲状腺无细胞癌
Metastatic site
胸腔积液
Synonyms
KMHDASH2, KMH2

Documents

KMH-2

Cell line · Product number 305142

KMH-2 is a human anaplastic thyroid carcinoma (ATC) cell line derived from a male patient with a rapidly progressing and fatal form of thyroid cancer. Ana...

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01General information

Description

KMH-2 是一种人类无细胞甲状腺癌(ATC)细胞系,来源于一名患有进展迅速且致命的甲状腺癌的男性患者。甲状腺无节细胞癌是最具侵袭性和致命性的甲状腺恶性肿瘤之一,其特点是生长迅速,对传统疗法具有抗药性。KMH-2 细胞是在患者接受任何化疗或放疗之前从原发肿瘤的活检中提取的。这些细胞对于研究 ATC 的病理生理学以及测试新治疗药物的疗效具有重要意义。

KMH-2 细胞系在体外培养时呈现纺锤形形态,这是许多无性甲状腺癌细胞的典型特征。这些细胞对多种化疗药物(包括顺铂、多柔比星、依托泊苷和百普霉素)具有抗药性,这反映了治疗甲状腺癌的临床挑战。KMH-2细胞的耐药性被归因于多药耐药性相关蛋白(MRP)mRNA的表达,尽管它们不表达与P-糖蛋白相关的mdr-1和mdr-3 mRNA,这表明它们的耐药机制与P-糖蛋白无关。这种对化疗的耐药性使 KMH-2 成为研究替代治疗策略的重要模型。

在生长特性方面,KMH-2 细胞的倍增时间相对较长,其致瘤性已在使用无胸腺裸鼠的异种移植模型中得到证实。然而,这些细胞需要特定的条件才能在体内增殖,如使用小塑料板促进接种后的生长。KMH-2的染色体分析发现了多种异常,这是侵袭性癌症的常见特征,这进一步突出了它们在研究甲状腺无节细胞癌遗传基础方面的作用。

Tissue
甲状腺
Disease
甲状腺无细胞癌
Metastatic site
胸腔积液
Synonyms
  • KMHDASH2
  • KMH2

02Classification

Organism
Human
Sex
Male
Growth properties
Adherent
Biosafety level
1

03Characteristics

Morphology
纺锤形细胞与巨细胞
Doubling time
58 小时

04Identifiers

Citation
KMH-2(Cytion 产品编号 305142)
NCBI TaxID
9606
Cellosaurus
CVCL_S641

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
71 年
Ethnicity
亚洲