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K7M2 wt-Luc
Tissue
腹水
Disease
小鼠骨肉瘤
Metastatic site
肺部
Synonyms
K7M2-WT, K7M2

Documents

K7M2 wt-Luc

Cell line · Product number 305684

K7M2 wt-Luc is a bioluminescent derivative of the murine K7M2 osteosarcoma cell line, engineered to stably express a firefly luciferase reporter gene. The...

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01General information

Description

K7M2 wt-Luc 是小鼠 K7M2 骨肉瘤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 K7M2 细胞系是从 BALB/c 小鼠自发性骨肉瘤的腹水中建立的,其特征为成骨细胞样形态和较高的转移潜力,尤其是向肺部的转移。 K7M2 是 BALB/c 背景中为数不多的同基因骨肉瘤模型之一,因此成为研究骨肉瘤生物学、肿瘤与免疫相互作用以及在免疫功能正常的宿主中评估免疫治疗策略的宝贵工具。

K7M2 wt-Luc 中稳定的荧光素酶整合,使得在同基因 BALB/c 小鼠中对原发肿瘤的生长和肺转移扩散进行灵敏、无创的生物发光成像 (BLI) 成为可能。 在给药荧光素底物后,发出的信号与存活肿瘤细胞数量呈正相关,从而支持对肿瘤植入、转移定植和治疗反应进行纵向监测,而无需在每个时间点进行侵入性操作。 这使得 K7M2 wt-Luc 特别适合用于评估骨肉瘤中抗肿瘤和抗转移药物、检查点抑制剂以及联合治疗策略的临床前研究。

K7M2 wt-Luc 保留了亲本 K7M2 系的生物学和免疫学特性,包括补体成分 C3 和 Fc γ 受体 I (FcgRI) 的表达,这些特征可能会影响与先天免疫系统的相互作用。该荧光素酶报告基因大大提高了实验的灵活性和灵敏度,可实时追踪肿瘤进展。 研究人员在进行大规模体内实验前,应根据其具体的实验条件,对荧光素酶活性、生长动力学及转移行为进行验证。

Tissue
腹水
Disease
小鼠骨肉瘤
Metastatic site
肺部
Applications
骨肉瘤研究;同基因型BALB/c肿瘤模型;基于生物发光技术的肺转移成像;抗肿瘤和抗转移药物评价;免疫检查点抑制剂检测;先天免疫相互作用研究(C3、FcgRI);小儿骨癌免疫治疗
Synonyms
  • K7M2-WT
  • K7M2

02Classification

Organism
Mouse
Sex
Female
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

Cell type
成骨细胞
Tumourigenic
是

04Identifiers

Citation
K7M2 wt-Luc(Cytion 产品编号 305684)
NCBI TaxID
10090

05Expression and susceptibility

Receptors expressed
补体(C3),表达,Fc 受体,IgG,高亲和力 I(Fcgr1),表达
Antigen expression
Luc2(萤火虫,密码子优化版)

06Handling 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。
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

Breed / subspecies
BALB/c
Donor age
895 天