BioVector® ROSA KIT D816V Luci / Gluc 人肥大细胞株 / ROSA KIT D816V Luci (Gluc) Human Mast Cell Line

一、 背景与来源说明 / Background & Origin

ROSA KIT D816V Luci（在文献中亦称为 ROSA KIT D816V-Gluc）是以 ROSA KIT D816V 母本细胞为基础，通过慢病毒介导转入高灵敏度高亮发光酶（如分泌型 Gaussia princeps 荧光素酶, Gluc / Firefly Luciferase）标记基因建立的报告细胞株。该细胞系保留了驱动型 KIT D816V 活化突变和肥大细胞功能受体。通过表达荧光素酶，该细胞株允许研究人员在无需杀死动物的情况下，通过无创外周血生物发光检测或小动物活体成像系统（IVIS）实时、定量监控小鼠异种移植模型（In Vivo Xenograft）中肥大细胞白血病（MCL）和系统性肥大细胞增多症（SM）的肿瘤负荷及药物疗效。

二、 细胞生物学特征 / Biological Characteristics

• 组织来源 / Tissue Origin：人造血祖细胞源肥大细胞 (Human Mast Cell Line)

• 生长特性 / Growth Properties：主要呈悬浮生长（Suspension）或部分弱贴壁生长

• 关键基因型 / Key Genotype：

  • KIT 外显子 17 D816V 激活型突变 (KIT D816V)

  • 稳定表达发光标记：Luciferase (Gluc / Luc) 与荧光蛋白（如 CFP/GFP）

• 细胞标记物 / Cell Markers：CD117 (KIT)、Fc epsilon RI (IgE 高亲和力受体)、Tryptase

• 应用领域 / Applications：活体动物模型（In Vivo）肿瘤负荷定量、抗 KIT D816V 靶向药物（如 Avapritinib、Midostaurin）体内药效评估、小动物高通量筛选

三、 培养条件与传代指南 / Culture & Handling Protocols

1. 完全培养基配方 / Medium Formulation

• 基础培养基 / Basal Medium：IMDM 或 RPMI-1640 基础培养基

• 血清与添加剂 / Serum & Supplements：

  • 10% 胎牛血清 (FBS)

  • 人干细胞因子 (rhSCF, 10 至 50 ng/mL，依特定株系对 SCF 依赖性而定)

  • 维持筛选抗生素（若有，如适量 Puromycin，根据克隆抗性配置）

• 抗生素 / Antibiotics：100 U/mL 青霉素 + 100 ug/mL 链霉素

2. 培养与传代操作 / Incubation & Subculture

• 培养环境 / Conditions：37 摄氏度，5% CO2，饱和湿度

• 密度管理 / Confluence：保持细胞密度在 2 x 10^5 至 1 x 10^6 cells/mL 之间，防止发光表达衰减或细胞生长停滞

• 传代步骤 / Procedure：

  1. 收集含有细胞的培养悬液于离心管中。

  2. 以 200 x g 离心 5 分钟，弃去旧培养基。

  3. 加入新鲜完全培养基重悬细胞沉淀。

  4. 按照 1:2 至 1:4 的比例稀释接种至新培养瓶中。

四、 技术指标简表 / Technical Data Summary

BioVector® ROSA KIT D816V Luci / Gluc Human Mast Cell Line

1. Background and Origin

ROSA KIT D816V Luci (also reported as ROSA KIT D816V-Gluc) was established by transducing parental ROSA KIT D816V human mast cells with a lentiviral vector expressing a bright luciferase reporter gene (such as secreted Gaussia princeps luciferase, Gluc, or Firefly Luciferase).Maintaining the driver KIT D816V activation mutation and characteristic mast cell surface markers, this cell line enables researchers to perform non-invasive bioluminescent imaging (IVIS) or quantitative blood luciferase detection to track tumor burden, disease kinetics, and drug efficacy in vivo using immunocompromised xenograft mouse models.

2. Biological Characteristics

• Tissue Origin: Human umbilical cord blood CD34+ derived mast cell line

• Growth Properties: Suspension or loosely adherent round cells

• Key Genotype:

  • KIT Exon 17 D816V gain-of-function mutation

  • Stable integration of Luciferase (Gluc / Luc) reporter

• Cell Markers:

  • Receptors: CD117 (KIT), Fc epsilon RI (IgE receptor)

  • Granule content: Mast cell Tryptase

• Applications: In vivo bioluminescence imaging (IVIS), systemic mastocytosis xenograft models, in vivo TKI efficacy assays, and tumor kinetics studies

3. Culture and Handling Protocols

Culture Medium Formulation

• Basal Medium: IMDM or RPMI-1640

• Serum and Additives: 10 percent Fetal Bovine Serum (FBS), supplemented with rhSCF (10 to 50 ng/mL) and selective antibiotics if needed

• Antibiotics: 100 U/mL Penicillin and 100 ug/mL Streptomycin

Incubation and Subculture Guide

• Incubation Conditions: 37 degrees Celsius, 5 percent CO2, humidified atmosphere

• Density Management: Maintain cell density between 2 x 10^5 and 1 x 10^6 cells/mL to preserve exponential growth and strong reporter expression

• Subculture Procedure:

  1. Collect cell suspension into a sterile centrifuge tube.

  2. Centrifuge at 200 x g for 5 minutes and remove supernatant.

  3. Resuspend cell pellet in fresh complete medium.

  4. Dilute and seed cells at a ratio of 1:2 to 1:4 into fresh culture vessels.

4. Technical Data Summary

BioVector NTCC质粒载体菌株细胞蛋白抗体基因保藏中心

电话:400-800-2947

工作QQ/微信同号:1843439339

E-mail:BioVector@163.com

网址http://www.biovector.net