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CDNR4
Tissue
乳房
Disease
腺癌

Documents

CDNR4

Cell line · Product number 400391

The CDNR4 cell line comprises a specialized subset originating from the COMMA-D cell line, known for modeling mouse mammary carcinoma. This clonal subpopu...

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1

01General information

Description

CDNR4 细胞系由源自 COMMA-D 细胞系的特异性亚群组成,该细胞系以模拟小鼠乳腺癌而闻名。对这一克隆亚群进行了广泛的表征,揭示了一系列独特的特性和功能。CDNR4 细胞最显著的特征之一是与乳腺干细胞相似,这使它们成为探索干细胞生物学、癌变和群体内细胞异质性等方面的重要资源。这些细胞是通过转染带有卡那霉素和新霉素抗性基因(Tn5基因)的转座子培育出来的,从而产生了各种有趣的特性和能力,包括分化成肿瘤前和肿瘤表型的潜力。

CDNR4 起源于 COMMA-D 株系,该株系最初利用相衬显微镜、免疫细胞化学染色、DNA 含量分析和致癌潜力评估等多种技术对其细胞异质性进行了研究,CDNR4 是一个与众不同的克隆。通过特定的转染和选择方法,像 CDNR4 这样的克隆亚群被分离出来,每个亚群都在一定程度上保持了原始 COMMA-D 亲本细胞的异质性。这种异质性的保持强调了这些细胞群的复杂性,提高了 CDNR4 细胞在细胞分化和癌症进展研究中的价值。

Tissue
乳房
Disease
腺癌

02Classification

Organism
Mouse
Sex
Female
Growth properties
Adherent
Biosafety level
1

03Identifiers

Citation
CDNR4(Cytion 目录编号 400391)
NCBI TaxID
10090
Cellosaurus
CVCL_5719

04Handling and culture

Culture medium
DMEM,w:4.5 克/升葡萄糖,w:4 毫摩尔 L-谷氨酰胺,w:3.7 克/升 NaHCO3,w:1.0 毫摩尔丙酮酸钠(Cytion 编号 820300a)。
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。
Seeding density
建议使用2×10⁴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.

05Quality 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。

06Donor

Donor age
1 年