BioVector? IMS-M2 人急性髓系白血病細胞株

BioVector? IMS-M2 Human Acute Myeloid Leukemia Cell Line

第一部分 中文說明

一 產(chǎn)品基本信息與遺傳學背景

• 細胞名稱：BioVector? IMS-M2 人急性髓系白血病細胞

• 系統(tǒng)學 Accession：Cellosaurus CVCL_RL93

• 物種來源：人類 (Homo sapiens)

• 性別與年齡：女性，59歲（Japanese 種群背景）

• 組織與疾病背景：該細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分型為 M2 型）的59歲女性患者的骨髓（Bone marrow）組織。

• 核心致癌分子與突變特征：

  • NPM1 基因第12外顯子突變（NPM1c+）：該細胞株雜合表達標志性的 NPM1 基因 A型突變（p.Trp288Cysfs*12）。NPM1 突變導致其編碼的核磷蛋白發(fā)生移碼，錯誤地定位積聚于細胞質(zhì)中（NPM1 胞質(zhì)異位），這是急性髓系白血病中最常見且極其核心的分子發(fā)病機制之一。

  • ETV6-NTRK3 融合基因（TEL-TRKC）：含有染色體隱匿性易位生成的 ETV6-NTRK3 原癌基因融合，該融合蛋白具有持續(xù)激活的酪氨酸激酶活性，可協(xié)同刺激細胞發(fā)生惡性增殖。

  • 復雜染色體核型變異：核型分析通常表現(xiàn)為 48, XX, add(6)(q27), +8, inv(12)(p13q15), +add(15)(q25) 等多倍體與結(jié)構(gòu)重排。

• 生物安全級別：1級（BSL-1）。

二 細胞形態(tài)學與培養(yǎng)環(huán)境

• 形態(tài)學特征：展現(xiàn)典型的原始白血病髓系細胞形態(tài)。鏡下觀察細胞呈圓形或卵圓形、胞質(zhì)邊界清晰，在培養(yǎng)液中以分散懸浮的單細胞形式生長。

• 生長模式：懸浮生長。

• 生長密度依耐性：該細胞對低接種濃度極其敏感。當細胞濃度過低時（如低于每毫升 $1.0 \times 10^5$ 個活細胞），細胞會由于缺乏自分泌生長因子的支持而出現(xiàn)增殖停滯、甚至發(fā)生大面積凋亡。

• 標準完全培養(yǎng)基配方：

  • 基礎培養(yǎng)基：BioVector? RPMI-1640 培養(yǎng)基。

  • 維持添加：

    • 10% 到 20% BioVector? 優(yōu)質(zhì)熱滅活胎牛血清（優(yōu)質(zhì)高濃度血清能顯著提升復蘇早期細胞的活率）。

    • 1% Penicillin-Streptomycin 雙抗溶液。

• 物理培養(yǎng)參數(shù)：37攝氏度恒溫、5% 二氧化碳、空氣飽和濕度。

三 細胞傳代與復蘇標準操作步驟

1. 常規(guī)傳代操作：

   • 核心維持密度：日常培養(yǎng)中務必將細胞密度嚴格控制在 每毫升 $2.0 \times 10^5$ 至 $2.0 \times 10^6$ 個活細胞 之間。

   • 當細胞密度達到或接近每毫升 $2.0 \times 10^6$ 個時，需要進行傳代。由于是完全懸浮生長，無需使用胰酶消化。

   • 將細胞懸液轉(zhuǎn)移至無菌離心管中，以 150 g 離心 5 分鐘，棄去舊培養(yǎng)基。

   • 傳代接種策略：加入新鮮預熱的完全培養(yǎng)基重懸，推薦起始接種密度切勿低于每毫升 $2.5 \times 10^5$ 個活細胞。常規(guī)傳代比例為 1比2 至 1比3，每 2 到 3 天需要補液或傳代一次。

2. 凍存細胞復蘇：

   • 從液氮中取出冷凍管，立即投入 37攝氏度 BioVector? 水浴鍋中持續(xù)高頻搖動使其快速融化，控制在 1 到 2 分鐘內(nèi)。

   • 迅速移入含有 5 到 7 mL 預熱完全培養(yǎng)基的無菌試管中，150 g 離心 5 分鐘以徹底去除殘留的 DMSO 凍存液。

   • 棄去上清，加入 4 到 5 mL 新鮮的高血清完全培養(yǎng)基重懸，接種至小培養(yǎng)瓶中，保持相對較高的初始復蘇細胞密度。

四 核心科研應用方向

1. NPM1 突變型白血病的靶向藥物篩查與致病機制解析：BioVector? IMS-M2 是國際上公認用于研究 NPM1 突變（NPM1c）急性髓系白血病非常珍貴的標準工具細胞。常被用于測試和開發(fā)各種新型的 NPM1 胞質(zhì)異位阻斷劑、NPM1 降解劑、以及天然多酚類化合物（如表沒食子兒茶素沒食子酸酯 EGCG）對 NPM1 表達下調(diào)、誘導白血病細胞發(fā)生凋亡與自噬的藥效動力學評價。

2. ETV6-NTRK3 融合激酶抑制劑（TRK Inhibitors）篩選：由于該細胞高表達具有持續(xù)酪氨酸激酶活性的 ETV6-NTRK3 融合基因，它是評估第一代及第二代廣譜 TRK 抑制劑（如 Larotrectinib, Entrectinib）在髓系白血病遺傳背景下抗增殖活性的靶向白血病細胞模型。

3. 髓系白血病細胞分化阻斷與表觀遺傳學調(diào)控研究：常用于通過誘導劑（如全反式維甲酸 ATRA、維生素D3 或 TPA）干預，觀察 IMS-M2 細胞表面分化抗原（如 CD13, CD15, CD33, HLA-DR）的表達豐度切換，深入解析 AML-M2 型白血病發(fā)病過程中髓系分化停滯的表觀遺傳阻斷位點。

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

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