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Lovo-Luc
Tissue
转移性
Disease
结肠腺癌
Metastatic site
左锁骨上淋巴结

Documents

Lovo-Luc

Cell line · Product number 305682

LoVo-Luc is a bioluminescent derivative of the human LoVo colorectal adenocarcinoma cell line, engineered to stably express a firefly luciferase reporter ...

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01General information

Description

LoVo-Luc 是人类 LoVo 结直肠腺癌细胞系的生物发光衍生物,经过基因工程改造,可稳定表达萤火虫荧光素酶报告基因。 亲本 LoVo 细胞系源自一名 56 岁男性患者结直肠腺癌的左侧锁骨上淋巴结转移灶,是癌症研究中应用最广泛的结直肠癌细胞系之一。 LoVo 细胞表现出上皮样形态,其特征为微卫星不稳定性 (MSI)、错配修复缺陷 (dMMR) 以及表达包括 Myc、myb、ras、fos 和 p53 在内的致癌基因。 这些特征使LoVo成为研究错配修复缺陷型结直肠癌生物学特性及治疗反应(包括对免疫治疗的敏感性)的理想模型。

LoVo-Luc中稳定的荧光素酶整合,使得利用免疫缺陷宿主进行异种移植模型中的肿瘤负荷敏感、定量生物发光成像(BLI)成为可能。发光信号与存活肿瘤细胞数量相关,支持对肿瘤植入、生长动力学及治疗反应进行无创纵向监测。 LoVo-Luc 在针对 MMR 缺陷型结直肠癌评估免疫检查点抑制剂、化疗药物和靶向治疗的临床前研究中具有特别重要的价值,同时也适用于高通量药物筛选以及结直肠癌生物学机制的研究。

LoVo-Luc 保留了亲本 LoVo 细胞系的已知分子特征,包括其微卫星不稳定性(MSI)表型、致癌基因表达谱以及癌胚抗原(CEA)的产生。荧光素酶报告基因显著提高了实验通量,并能够对治疗疗效进行实时药效学评估。 研究人员在将其用于大规模临床前研究之前,应根据具体实验条件验证荧光素酶活性、MMR状态及细胞生长动力学。

Tissue
转移性
Disease
结肠腺癌
Metastatic site
左锁骨上淋巴结

02Classification

Organism
Human
Sex
Male
Growth properties
Adherent
GMO status
GMO-S1
GMO status details
GMO-S1: This cell line contains a stably integrated firefly luciferase reporter cassette (Luc2, codon-optimized) introduced via replication-incompetent lentiviral transduction. The resulting polyclonal cell population was maintained under puromycin selection (1–5 µg/mL). S1 containment is required. This classification applies only within Germany and may differ elsewhere.
Biosafety level
1

03Characteristics

Morphology
上皮样
Tumourigenic
是,在裸鼠身上
Doubling time
24-48小时

04Identifiers

Citation
Lovo-Luc(Cytion 产品编号 305682)
NCBI TaxID
9606
Cellosaurus
CVCL_4Y03

05Expression and susceptibility

Antigen expression
HLA A11、B15、B17、Cw1、Cw3、B型血、Luc2(萤火虫,密码子优化)
Isoenzymes
G6PD,B,PGM1,2,PGM3,1-2,6PGD,A,ES-D,1
Oncogenes
Myc +, myb + , ras +, fos +, p53 +, sis -, abl -, ros -, src -
Mutational profile
突变:p.Lys437Argfs*5,杂合子;突变:p.Arg1114Ter,杂合子;突变:p.Met1431fs*42,杂合子; 突变:p.Arg2816Gln,杂合子;突变:p.Leu15Phefs*41,杂合子;突变:p.Arg505Cys,杂合子; 突变:p.Gly13Asp,杂合子;突变:p.Ala292Val,杂合子;突变:p.Lys128Serfs*35,纯合子
Reverse transcriptase
阴性
Products
癌胚抗原 (CEA) 908 纳克/106 个细胞/10 天

06Handling and culture

Culture medium
Ham's F12K 培养基,w:2.0 mM L-谷氨酰胺,w:2.0 mM 丙酮酸钠,w:2.5 g/L NaHCO3(Cytion 文章编号 820608a)。
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。
Split ratio
1 至 3
Seeding density
1-3×10⁴/cm²
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 200 x g for 5 minutes, carefully discard the supernatant containing freezing medium.

  7. Follow the procedure described under Post-Thaw Recovery

07Quality control and cryopreservation

Freeze medium
作为冷冻保存培养基,我们使用完全生长培养基+10% DMSO,以获得足够的解冻后存活率。
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

Donor age
56 岁
Ethnicity
高加索人