BioVector® MB-02 细胞株（人急性巨核细胞白血病细胞） / BioVector® MB-02 Cell Line (Human Acute Megakaryoblastic Leukemia Cell Line)

一、 背景与来源说明 / Background & Origin

BioVector® MB-02 是一种来源于一名患有急性巨核细胞白血病（Acute Megakaryoblastic Leukemia, AMKL / FAB-M7）患者骨髓的人源巨核/红系祖细胞株（Human Megakaryocytic/Erythroid Progenitor Cell Line）。

MB-02 细胞株是研究巨核细胞分化（Megakaryopoiesis）、红系细胞分化（Erythropoiesis）以及细胞因子依赖性造血信号传导的经典体外模型。该细胞株在基础状态下高度依赖促红细胞生成素（EPO）或促血小板生成素（TPO）/ GM-CSF 生长，表达巨核系与红系双重表面标记。在 EPO 或 PMA（佛波酯）诱导下，MB-02 细胞可表现出向成熟巨核细胞（表达 CD41/CD61 并出现多倍体化）或红系细胞（表达血红蛋白）双向分化的潜力，广泛应用于急性巨核细胞白血病发病机制、造血干细胞分化调控及抗白血病药物筛选。

二、 细胞生物学特征 / Biological Characteristics

• 物种与来源 / Taxonomy & Source：人（Homo sapiens），骨髓（急性巨核细胞白血病 AMKL / M7）

• 生物安全等级 / Biosafety Level：BSL-1（或根据人源细胞规范按 BSL-2 防护）

• 生长特性 / Growth Properties：悬浮生长（Suspension），呈圆形、单核或多核巨核前体细胞形态

• 细胞因子依赖性 / Cytokine Dependency：重度依赖 EPO (Erythropoietin) 或 GM-CSF / TPO 维持持续增殖与生存

• 表面表型与标记物 / Surface Markers & Expression：

  • 巨核系标记：CD41a (GPIIb)(+)、CD61 (GPIIIIa)(+)、CD42b (GPIb)(+)

  • 红系及祖细胞标记：Glycophorin A (CD235a)(+)、CD34(+)、CD117 (c-Kit)(+)

• 主要应用 / Key Applications：巨核细胞与红系双向分化机制研究、EPO/TPO 信号转导分析、AMKL（M7 型白血病）靶向药物筛选、多倍体形成与血小板生成研究

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

1. 推荐培养基 / Complete Growth Medium

• 基础培养基：BioVector® RPMI-1640 Medium 或 BioVector® IMDM Medium

• 添加成分：10%–15% 灭活 BioVector® Fetal Bovine Serum (FBS) + 1% 双抗（Penicillin-Streptomycin）

• 必需细胞因子 / Vital Supplement：必需添加重组人促红细胞生成素 rhEPO (1–2 U/mL) 或 rhGM-CSF (10 ng/mL) / rhTPO (10–20 ng/mL)，缺乏细胞因子将迅速导致细胞凋亡

• 培养环境：37 摄氏度，5% $\text{CO}_2$，高湿度培养箱

2. 复苏与传代操作 / Thawing & Subculture

1. 复苏：将冻存管快速置于 37 摄氏度水浴中融化（1–2 分钟）。加入 8–10 mL 预热并补充有 rhEPO 的 BioVector® Complete Medium，120 x g 离心 5 分钟，弃上清后重悬接种于培养瓶中。

2. 传代与密度控制：

   • 细胞密度：维持密度在 $2 \times 10^5 \text{ cells/mL}$ 至 $1 \times 10^6 \text{ cells/mL}$ 之间。

   • 换液传代：每 2–3 天通过离心（120 x g, 5 分钟）更换新鲜培养基，并补足 rhEPO。无需 Trypsin 消化。

3. 冻存保存 / Cryopreservation

• 冻存液配方：90% BioVector® FBS + 10% DMSO（包含 2 U/mL rhEPO）

• 保存条件：程序降温后保存于液氮（-196 摄氏度）。

四、 技术指标简表 / Technical Data Summary

BioVector® MB-02 细胞株（人急性巨核细胞白血病细胞） / BioVector® MB-02 Cell Line (Human Acute Megakaryoblastic Leukemia Cell Line)

1. Background and Origin

BioVector® MB-02 is a human megakaryocytic/erythroid progenitor cell line established from the bone marrow of a patient with acute megakaryoblastic leukemia (AMKL / FAB-M7).

The MB-02 cell line is a standard human model for investigating megakaryopoiesis, erythropoiesis, and cytokine-dependent signal transduction. MB-02 cells are strictly dependent on erythropoietin (EPO) or TPO / GM-CSF for survival and proliferation. Upon stimulation with specific lineage inducers (such as EPO, TPO, or PMA), MB-02 cells can undergo bipotential differentiation into mature megakaryocytes (expressing CD41/CD61 with polyploidization) or erythroid lineage cells, making it an essential platform for megakaryoblastic leukemia research and therapeutic screening.

2. Biological Characteristics

• Taxonomy & Source: Homo sapiens, Bone marrow (Acute Megakaryoblastic Leukemia, AMKL)

• Biosafety Level: BSL-1 (or BSL-2 per institutional biosafety guidelines)

• Growth Properties: Suspension; displays rounded progenitor cell morphology

• Cytokine Dependency: Strictly dependent on EPO (Erythropoietin) or GM-CSF/TPO for proliferation

• Cell Surface Markers:

  • Megakaryocytic Markers: CD41a(+), CD61(+), CD42b(+)

  • Erythroid & Stem Markers: Glycophorin A (CD235a)(+), CD34(+), CD117 (c-Kit)(+)

• Primary Applications: Megakaryocyte and erythroid differentiation, EPO/TPO signaling mechanisms, AMKL pathogenesis, and high-throughput drug screening

3. Culture and Handling Protocols

Recommended Growth Medium

• Basal Medium: BioVector® RPMI-1640 Medium or BioVector® IMDM Medium

• Supplements: 10%–15% heat-inactivated BioVector® Fetal Bovine Serum (FBS) + 1% Penicillin-Streptomycin

• Required Cytokine: Recombinant human EPO (rhEPO, 1–2 U/mL) or rhGM-CSF (10 ng/mL) / rhTPO (10–20 ng/mL). Omission of cytokines leads to rapid apoptotic cell death.

• Culture Conditions: 37 degrees Celsius, 5% $\text{CO}_2$, humidified incubator

Thawing and Subculture Protocol

1. Thawing: Thaw rapidly in a 37 degrees Celsius water bath. Dilute in pre-warmed BioVector® Complete Medium containing rhEPO, centrifuge at 120 x g for 5 minutes, and seed into culture flasks.

2. Passaging:

   • Maintain cell density between $2 \times 10^5 \text{ cells/mL}$ and $1 \times 10^6 \text{ cells/mL}$.

   • Subculture every 2–3 days by centrifugation (120 x g, 5 minutes) and resuspension in fresh cytokine-supplemented medium.

Cryopreservation

• Freezing Medium: 90% BioVector® FBS + 10% DMSO + 2 U/mL rhEPO

• Storage: Liquid nitrogen (-196 degrees Celsius).

4. Technical Data Summary

BioVector NTCC质粒载体菌株细胞蛋白抗体基因保藏中心

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