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CT26.CL25
Tissue
结肠
Disease
腺癌
Synonyms
CT26-clone 25

Documents

CT26.CL25

Cell line · Product number 305353

The CT26.CL25 cell line is a murine colon carcinoma model derived from the parental CT26 cell line, which is a chemically induced, undifferentiated colon ...

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  • CoA
1

01General information

Description

CT26.CL25细胞系是一种小鼠结肠癌模型,来源于亲代CT26细胞系,后者是一种化学诱导的未分化结肠癌,来源于BALB/c小鼠。CT26.CL25经过基因改造,能表达β-半乳糖苷酶(β-gal)蛋白,是研究肿瘤免疫学和免疫疗法,特别是肿瘤相关抗原(TAAs)的绝佳模型。通过这种改造,可以针对作为新抗原的β-gal进行特异性免疫学研究,从而促进肿瘤免疫逃避机制的研究以及癌症疫苗或采用性细胞疗法的开发。

CT26.CL25 已被用于临床前模型,以研究免疫反应和免疫疗法的疗效,如使用负载有肿瘤相关抗原的树突状细胞 (DC)。研究表明,使用从逆转录病毒抗原(如 gp70)中提取的多肽脉冲 DC 的免疫策略可引起强有力的抗肿瘤免疫反应。在实验模型中,观察到了针对 gp70 的 CD8+ 细胞毒性 T 淋巴细胞(CTL)的活化,这证明了细胞系在测试免疫治疗方法中的实用性。然而,用这种多肽载荷的 DCs 进行免疫有其局限性,特别是在治疗已确诊的转移瘤方面,这凸显了将预防性免疫反应转化为治疗效果所面临的挑战。

此外,CT26.CL25 经常被用于测试联合免疫疗法的疗效,如使用免疫检查点抑制剂或癌症疫苗。例如,有研究评估了节律化疗联合免疫检查点抑制剂的影响,其中 CT26.CL25 诱导免疫原性细胞死亡(ICD)对增强抗肿瘤免疫反应至关重要。这些研究表明,以免疫检查点为靶点可与化疗协同提高肿瘤排斥率并建立长期免疫记忆。

Tissue
结肠
Disease
腺癌
Synonyms
  • CT26-clone 25

02Classification

Organism
Mouse
Sex
Female
Growth properties
Adherent
GMO status
GMO-S1
GMO status details
GMO-S1: This murine colon carcinoma cell line (CT26.CL25) contains a retroviral vector encoding lacZ and Tn5-neo, enabling β-galactosidase expression and neomycin resistance. The construct is stably integrated into CT26 cells. This classification applies only within Germany and may differ elsewhere.
Biosafety level
1

03Characteristics

Morphology
成纤维细胞
Tumourigenic
是,在 BALB/c 小鼠中

04Identifiers

Citation
CT26.CL25(Cytion 目录号 305353)
NCBI TaxID
10090
Cellosaurus
CVCL_7255

05Expression and susceptibility

Antigen expression
H-2d
Mutational profile
基因缺失:基因缺失:Cdkn2a,同型;突变:Kras,p.Gly12Asp(c.35G>A),同型:Kras,p.Gly12Asp (c.35G>A),同型
Products
表达的基因:β-半乳糖苷酶(β-gal)、H-2D

06Handling 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、1% NEAA、0.4 mg/mL G418、2.5 g/L 葡萄糖和 10 mM HEPES。
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.

07Quality 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。

08Donor

Breed / subspecies
BALB/c
Donor age
未说明