BioVector® IMS-M2 人急性髓系白血病细胞株

BioVector® IMS-M2 Human Acute Myeloid Leukemia Cell Line

第一部分 中文说明

一 产品基本信息与遗传学背景

• 细胞名称：BioVector® IMS-M2 人急性髓系白血病细胞

• 系统学 Accession：Cellosaurus CVCL_RL93

• 物种来源：人类 (Homo sapiens)

• 性别与年龄：女性，59岁（Japanese 种群背景）

• 组织与疾病背景：该细胞株建立于一名患有急性髓系白血病部分成熟型（Acute Myeloid Leukemia, FAB分型为 M2 型）的59岁女性患者的骨髓（Bone marrow）组织。

• 核心致癌分子与突变特征：

  • NPM1 基因第12外显子突变（NPM1c+）：该细胞株杂合表达标志性的 NPM1 基因 A型突变（p.Trp288Cysfs*12）。NPM1 突变导致其编码的核磷蛋白发生移码，错误地定位积聚于细胞质中（NPM1 胞质异位），这是急性髓系白血病中最常见且极其核心的分子发病机制之一。

  • ETV6-NTRK3 融合基因（TEL-TRKC）：含有染色体隐匿性易位生成的 ETV6-NTRK3 原癌基因融合，该融合蛋白具有持续激活的酪氨酸激酶活性，可协同刺激细胞发生恶性增殖。

  • 复杂染色体核型变异：核型分析通常表现为 48, XX, add(6)(q27), +8, inv(12)(p13q15), +add(15)(q25) 等多倍体与结构重排。

• 生物安全级别：1级（BSL-1）。

二 细胞形态学与培养环境

• 形态学特征：展现典型的原始白血病髓系细胞形态。镜下观察细胞呈圆形或卵圆形、胞质边界清晰，在培养液中以分散悬浮的单细胞形式生长。

• 生长模式：悬浮生长。

• 生长密度依耐性：该细胞对低接种浓度极其敏感。当细胞浓度过低时（如低于每毫升 $1.0 \times 10^5$ 个活细胞），细胞会由于缺乏自分泌生长因子的支持而出现增殖停滞、甚至发生大面积凋亡。

• 标准完全培养基配方：

  • 基础培养基：BioVector® RPMI-1640 培养基。

  • 维持添加：

    • 10% 到 20% BioVector® 优质热灭活胎牛血清（优质高浓度血清能显著提升复苏早期细胞的活率）。

    • 1% Penicillin-Streptomycin 双抗溶液。

• 物理培养参数：37摄氏度恒温、5% 二氧化碳、空气饱和湿度。

三 细胞传代与复苏标准操作步骤

1. 常规传代操作：

   • 核心维持密度：日常培养中务必将细胞密度严格控制在 每毫升 $2.0 \times 10^5$ 至 $2.0 \times 10^6$ 个活细胞 之间。

   • 当细胞密度达到或接近每毫升 $2.0 \times 10^6$ 个时，需要进行传代。由于是完全悬浮生长，无需使用胰酶消化。

   • 将细胞悬液转移至无菌离心管中，以 150 g 离心 5 分钟，弃去旧培养基。

   • 传代接种策略：加入新鲜预热的完全培养基重悬，推荐起始接种密度切勿低于每毫升 $2.5 \times 10^5$ 个活细胞。常规传代比例为 1比2 至 1比3，每 2 到 3 天需要补液或传代一次。

2. 冻存细胞复苏：

   • 从液氮中取出冷冻管，立即投入 37摄氏度 BioVector® 水浴锅中持续高频摇动使其快速融化，控制在 1 到 2 分钟内。

   • 迅速移入含有 5 到 7 mL 预热完全培养基的无菌试管中，150 g 离心 5 分钟以彻底去除残留的 DMSO 冻存液。

   • 弃去上清，加入 4 到 5 mL 新鲜的高血清完全培养基重悬，接种至小培养瓶中，保持相对较高的初始复苏细胞密度。

四 核心科研应用方向

1. NPM1 突变型白血病的靶向药物筛查与致病机制解析：BioVector® IMS-M2 是国际上公认用于研究 NPM1 突变（NPM1c）急性髓系白血病非常珍贵的标准工具细胞。常被用于测试和开发各种新型的 NPM1 胞质异位阻断剂、NPM1 降解剂、以及天然多酚类化合物（如表没食子儿茶素没食子酸酯 EGCG）对 NPM1 表达下调、诱导白血病细胞发生凋亡与自噬的药效动力学评价。

2. ETV6-NTRK3 融合激酶抑制剂（TRK Inhibitors）筛选：由于该细胞高表达具有持续酪氨酸激酶活性的 ETV6-NTRK3 融合基因，它是评估第一代及第二代广谱 TRK 抑制剂（如 Larotrectinib, Entrectinib）在髓系白血病遗传背景下抗增殖活性的靶向白血病细胞模型。

3. 髓系白血病细胞分化阻断与表观遗传学调控研究：常用于通过诱导剂（如全反式维甲酸 ATRA、维生素D3 或 TPA）干预，观察 IMS-M2 细胞表面分化抗原（如 CD13, CD15, CD33, HLA-DR）的表达丰度切换，深入解析 AML-M2 型白血病发病过程中髓系分化停滞的表观遗传阻断位点。

PART 2 ENGLISH SECTION

I General Information and Genetic Background

• Cell Line Name: BioVector® IMS-M2

• Synonyms: IMSM2, IMS-M2

• Cellosaurus Accession: CVCL_RL93

• Species Origin: Human (Homo sapiens)

• Sex and Age of Donor: Female, 59 years old (Japanese population profile)

• Tissue and Disease Background: Established from the bone marrow aspirate of a 59-year-old female patient diagnosed with Acute Myeloid Leukemia with maturation (FAB classification: AML-M2 subtype).

• Core Oncogenic Markers & Genomic Traits:

  • NPM1 Exon 12 Mutation (NPM1c+): Heterozygously harbors the classic NPM1 Type A frame-shift mutation (p.Trp288Cysfs*12). This genomic lesion forces the nucleophosmin protein out of the nucleolus, accumulating abnormally inside the cytoplasm (cytoplasmic dislocation of NPM1), which represents one of the most prominent driver mechanisms in acute myeloid leukemia pathogenesis.

  • ETV6-NTRK3 Gene Fusion (TEL-TRKC): Carries the cryptic chromosomal translocation giving rise to the oncogenic ETV6-NTRK3 construct.The encoded chimeric protein functions as a constitutively active tyrosine kinase, synergistically driving leukemic survival pathways.

  • Complex Karyotype: Typically characterized by chromosomal abnormalities including a polyploid baseline of 48, XX, add(6)(q27), +8, inv(12)(p13q15), +add(15)(q25).

• Biosafety Level: BSL-1.

II Morphological Attributes and Cultivation Media

• Morphology: Displays classic primitive myeloblastic features. Under oil-immersion light microscopy, cells present round or oval profiles with distinct cytoplasmic boundaries, proliferating entirely as an unattached single-cell suspension.

• Growth Mode: Suspension growth.

• Density Dependency Alert: Highly vulnerable to low biomass initialization. If the cellular seed pool plummets below critical thresholds (e.g., $< 1.0 \times 10^5$ viable cells/mL), proliferation stalls completely due to a lack of autocrine growth factor reinforcement, frequently cascading into irreversible culture decay or apoptosis.

• Standard Complete Growth Medium Formulation:

  • Basal Medium: BioVector® RPMI-1640 medium.

  • Routine Supplements:

    • 10% to 20% premium BioVector® Heat-Inactivated Fetal Bovine Serum (FBS). High serum concentration safeguards structural recovery and viability post-thaw.

    • 1% Penicillin-Streptomycin solution.

• Physical Incubation Thresholds: Regulated strictly at 37 degrees Celsius under an atmospheric layer of 5% Carbon Dioxide and saturated air humidity.

III Subculturing and Thawing Protocols

1. Routine Passaging Schedule:

   • Core Density Window: Maintain the active suspension profile tightly bounded between $2.0 \times 10^5$ and $2.0 \times 10^6$ viable cells/mL.

   • Subculture the vessel when the active density reaches or approaches the $2.0 \times 10^6$ cells/mL baseline. Because the line expands purely in suspension, enzymatic trypsinization is unnecessary.

   • Pellet the slurry via centrifugation at 150 g for 5 minutes, and thoroughly decant the spent media.

   • Seeding Target: Resuspend the cells in fresh complete broth. Crucially, never re-seed the fresh vessel at densities lower than $2.5 \times 10^5$ viable cells/mL. The recommended split sequence averages 1:2 to 1:3, demanding culture splitting or feeding every 2 to 3 days.

2. Cryovial Thawing and Recovery:

   • Retrieve the cryovial from storage and submerge it into a 37 degrees Celsius BioVector® water bath with rapid agitation until completely liquefied within 1 to 2 minutes.

   • Promptly transfer the cell suspension into a sterile tube carrying 5 to 7 mL of pre-warmed complete growth medium and spin at 150 g for 5 minutes to fully purge residual DMSO.

   • Decant the supernatant, gently resuspend the cell pellet in fresh, serum-enriched complete BioVector® medium, and plate. Maintain a relatively high cell density during the immediate post-thaw cultivation phase to maximize viability.

IV Strategic Research Applications

1. NPM1-Mutated AML Targeted Therapy Screening and Epigenetics: BioVector® IMS-M2 stands as an excellent, internationally recognized reference model for evaluating NPM1-mutated (NPM1c) acute myeloid leukemia therapeutics. It is extensively utilized to screen small-molecule NPM1 cytoplasmic mislocalization blockers, targeted NPM1 degraders, and natural bioactive compounds (such as (-)-epigallocatechin-3-gallate, EGCG) to dissect down-regulation loops and apoptotic networks.

2. ETV6-NTRK3 Chimeric Kinase Profiling: Possessing the constitutively active ETV6-NTRK3 fusion architecture, this model serves as a specific assay platform to evaluate first- and second-generation pan-TRK small molecule tyrosine kinase inhibitors (e.g., Larotrectinib, Entrectinib) within a realistic myeloid malignancy setting.

3. Elucidating Myeloid Differentiation Blockades: Deployed alongside standard chemical differentiation vectors (such as All-Trans Retinoic Acid ATRA, Vitamin D3, or TPA) to track changes in surface immunophenotypic profiles (measuring variations in myeloid cluster antigens CD13, CD15, CD33, and HLA-DR), aiding the structural decryption of epigenetic blocks that arrest maturation in AML-M2 leukemia blasts.

d

BioVector NTCC质粒载体菌株细胞蛋白抗体基因保藏中心

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