BioVector® UKE-1 细胞株（人骨髓增殖性肿瘤/原发性血小板增多症细胞） / BioVector® UKE-1 Cell Line (Human Essential Thrombocythemia Cell Line)

一、 背景与来源说明 / Background & Origin

BioVector® UKE-1（人骨髓增殖性肿瘤/血小板增多症细胞株）是一种来源于一名患有原发性血小板增多症（Essential Thrombocythemia, ET）并随后转化为急性髓系白血病（Acute Myeloid Leukemia, AML）的患者外周血/骨髓的人源巨核细胞/髓系前体细胞株（Human Megakaryocytic/Myeloid Cell Line）。

UKE-1 细胞株是研究 JAK2 V617F 基因突变（该突变在骨髓增殖性肿瘤 MPNs 如真性红细胞增多症 PV、原发性血小板增多症 ET 及原发性骨髓纤维化 PMF 中高度高发）机制的经典细胞模型之一。UKE-1 细胞纯合表达 JAK2 V617F 突变基因，导致其下游 JAK-STAT、PI3K-AKT 及 MAPK/ERK 信号通路持续性异常激活。该细胞株广泛用于 MPNs 病理机制探究、JAK2 激酶小分子抑制剂（如 Ruxolitinib、Fedratinib）的筛选与耐药性评估以及巨核细胞系分化机制研究。

二、 细胞生物学特征 / Biological Characteristics

• 物种与来源 / Taxonomy & Source：人（Homo sapiens），外周血/骨髓（原发性血小板增多症/白血病）

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

• 生长特性 / Growth Properties：悬浮生长（Suspension）或部分松散贴壁（Loosely Adherent），呈圆形或多形性单核/多核巨核前体细胞形态

• 基因型特征 / Key Genetic Mutations：

  • JAK2 基因：纯合突变 JAK2 V617F (+/+)

  • TP53 基因：突变或缺失（导致细胞抗凋亡能力增强）

• 关键免疫表型 / Key Surface Markers：

  • 表达巨核/血小板系与髓系表面标记：CD33(+)、CD34(+)、CD41 (GPIIb)(+)、CD61 (GPIIIa)(+)、CD117 (c-Kit)(+)

• 主要应用 / Key Applications：骨髓增殖性肿瘤（MPNs）及 ET 病理机制研究、JAK2 抑制剂高通量筛选与靶向治疗、JAK-STAT 通路信号传导分析、巨核细胞分化与血小板生成机制

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® IMDM Medium 或 BioVector® RPMI-1640 Medium

• 添加成分：10%–15% 灭活 BioVector® Fetal Bovine Serum (FBS) + 10% 灭活马血清（Horse Serum，可选辅助成分）+ 1% 双抗（Penicillin-Streptomycin）

• 细胞因子（可选/视具体克隆而定）：部分亚株可在无外源细胞因子下生长，添加低浓度 TPO (Thrombopoietin) 或 GM-CSF 可促进巨核系分化

• 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 摄氏度水浴中摇晃融化（1–2 分钟）。加入 8–10 mL 预热的 BioVector® Complete Growth Medium，120 x g 离心 5 分钟，弃上清后用完全培养基重悬，置于培养瓶中悬浮培养。

2. 传代与密度控制：

   • 密度范围：维持细胞密度在 $3 \times 10^5 \text{ cells/mL}$ 至 $1.5 \times 10^6 \text{ cells/mL}$ 之间。

   • 传代方法：每 2–3 天离心（120 x g, 5 分钟）更换部分培养基，或直接加入新鲜 BioVector® Complete Medium 半量换液传代。无需使用 Trypsin 消化。

3. 冻存保存 / Cryopreservation

• 冻存液配方：90% BioVector® FBS + 10% DMSO 或 70% BioVector® Basal Medium + 20% BioVector® FBS + 10% DMSO

• 保存条件：程序降温后长期保存于液氮（-196 摄氏度）。

四、 技术指标简表 / Technical Data Summary

BioVector® UKE-1 细胞株（人骨髓增殖性肿瘤/原发性血小板增多症细胞） / BioVector® UKE-1 Cell Line (Human Essential Thrombocythemia Cell Line)

1. Background and Origin

BioVector® UKE-1 is a human megakaryocytic/myeloid leukemia cell line derived from the peripheral blood/bone marrow of a patient diagnosed with essential thrombocythemia (ET) that subsequently transformed into acute myeloid leukemia (AML).

The UKE-1 cell line is an established model for studying JAK2 V617F-mutated myeloproliferative neoplasms (MPNs). UKE-1 cells carry a homozygous JAK2 V617F mutation, resulting in constitutive hyperactivation of downstream signaling pathways, including JAK-STAT, PI3K-AKT, and MAPK/ERK. It serves as an essential tool for evaluating small-molecule JAK2 inhibitors (e.g., Ruxolitinib, Fedratinib), overcoming drug resistance, and dissecting megakaryocytic lineage differentiation.

2. Biological Characteristics

• Taxonomy & Source: Homo sapiens, Peripheral blood / Bone marrow (Essential Thrombocythemia / AML)

• Biosafety Level: BSL-1 (or BSL-2 per institutional guidelines for human cell lines)

• Growth Properties: Suspension or loosely adherent single/multinucleated megakaryocytic progenitor cells

• Genetic Features:

  • JAK2 Gene: Homozygous JAK2 V617F (+/+) mutation

  • TP53 Gene: Mutated / dysfunctional

• Key Surface Markers:

  • Expresses megakaryocytic and myeloid markers: CD33(+), CD34(+), CD41 (GPIIb)(+), CD61 (GPIIIIa)(+), CD117 (c-Kit)(+)

• Primary Applications: Pathogenesis of MPNs and essential thrombocythemia, screening of novel JAK2 inhibitors, JAK-STAT pathway signal transduction, and megakaryopoiesis studies

3. Culture and Handling Protocols

Recommended Growth Medium

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

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

• 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. Resuspend in 8–10 mL pre-warmed BioVector® Complete Growth Medium, centrifuge at 120 x g for 5 minutes, and seed into culture flasks.

2. Passaging:

   • Maintain cell density between $3 \times 10^5 \text{ cells/mL}$ and $1.5 \times 10^6 \text{ cells/mL}$.

   • Replenish or split culture every 2–3 days by centrifuging at 120 x g for 5 minutes or performing a partial medium dilution. No trypsinization is required.

Cryopreservation

• Freezing Medium: 90% BioVector® FBS + 10% DMSO or 70% BioVector® Basal Medium + 20% BioVector® FBS + 10% DMSO

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

4. Technical Data Summary

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