BioVector® N/TERT-1 人永生化角质形成细胞系 / BioVector® N/TERT-1 Human Immortalized Keratinocyte Cell Line

通用定义 / General Definition:BioVector® N/TERT-1 是一种广泛使用的人源永生化皮肤角质形成细胞系。该细胞系通过在原代人包皮角质形成细胞中稳定表达端粒酶逆转录酶 (hTERT) 并结合 p16INK4a 表达缺失而建立。与传统的通过病毒癌基因（如 HPV-16 E6/E7）转化的细胞系不同，N/TERT-1 在很大程度上保留了原代角质形成细胞的生物学特性，包括正常的核型、对分化信号的反应能力以及在器官型培养中形成分层表皮的能力。它被认为是研究皮肤生物学、毒理学和皮肤疾病体外建模的理想选择。

BioVector® N/TERT-1 is a widely used immortalized human skin keratinocyte cell line. It was established by stably expressing the human telomerase reverse transcriptase (hTERT) in primary human foreskin keratinocytes, coupled with the spontaneous loss of p16INK4a expression. Unlike traditional lines transformed by viral oncogenes (such as HPV-16 E6/E7), N/TERT-1 largely retains the biological characteristics of primary keratinocytes, including a normal karyotype, responsiveness to differentiation signals, and the ability to form a stratified epidermis in organotypic cultures. It is considered an ideal model for studying skin biology, toxicology, and in vitro modeling of dermatological diseases.

BioVector® N/TERT-1 技术说明书 (Technical Datasheet)

中文版说明书 (Chinese Datasheet)

1. 产品基本信息

• 产品名称： BioVector® N/TERT-1 人永生化角质形成细胞

• 组织来源： 人新生儿包皮组织

• 永生化机制： hTERT 稳定表达 + p16 缺失

• 生长特性： 贴壁生长

• 细胞形态： 典型的铺路石状上皮形态

2. 培养条件

• 基础培养基： BioVector® K-SFM（角质形成细胞无血清培养基），含牛垂体提取物 (BPE) 和表皮生长因子 (EGF)。

• 备选培养基：DFK 培养基（DMEM/F-12 1:1 混合，添加特定生长因子）。

• 钙浓度控制： 维持低钙浓度（通常小于 0.1 mM）以保持细胞处于未分化的增殖状态。

• 培养环境： 37 摄氏度，5% CO2。

• 传代比例： 1:3 至 1:6；当汇合度达到 70%-80% 时必须传代，以防止自发分化。

3. 细胞应用

• 皮肤替代模型： 用于构建 3D 有机型皮肤等效模型 (Organotypic Skin Equivalents, OSE)，研究表皮屏障功能。

• 毒理学测试： 评估化妆品、药物或化学物质对皮肤的刺激性和毒性。

• 炎症研究： 通过刺激（如 TNF-alpha）研究角质形成细胞的细胞因子分泌和信号通路。

4. 注意事项

• 密度敏感性： N/TERT-1 对细胞密度非常敏感。汇合度过高会触发钙依赖性分化过程，导致细胞停止增殖并脱落。

• 消化过程： 使用 0.05% Trypsin-EDTA 消化，一旦细胞变圆变亮即加入胰酶抑制剂或含血清培养基终止。

English Datasheet

1. General Product Information

• Product Name: BioVector® N/TERT-1 Human Immortalized Keratinocyte Cell Line

• Source: Human neonatal foreskin tissue

• Immortalization: hTERT stable expression + p16INK4a loss

• Growth Properties: Adherent

• Morphology: Classic cobblestone epithelial morphology

2. Culture Conditions

• Basal Medium: BioVector® K-SFM (Keratinocyte Serum-Free Medium) supplemented with Bovine Pituitary Extract (BPE) and Human Recombinant EGF.

• Alternative Medium:DFK medium (DMEM/F-12 mix with specific growth factors).

• Calcium Regulation: Maintain low calcium levels (typically < 0.1 mM) to keep cells in a proliferative, undifferentiated basal state.

• Incubation: 37°C, 5% CO2.

• Subculturing: 1:3 to 1:6 ratio; subculture at 70%-80% confluence to avoid density-induced differentiation.

3. Applications

• 3D Skin Modeling: Widely used in air-liquid interface (ALI) cultures to generate stratified human skin equivalents.

• Dermatotoxicity: Assessing the safety and irritancy of topical formulations or environmental pollutants.

• Gene Editing: An excellent platform for CRISPR/Cas9 or RNAi studies due to its consistent growth and primary-like genetic profile.

4. Key Usage Notes

• Differentiation Control: Because these cells retain the ability to differentiate, maintaining them in a sub-confluent state is critical for long-term culture stability.

• Coating: While they generally attach well to tissue-culture treated plastic, coating with Collagen I or IV can improve attachment and expansion in certain experimental setups.

注意 / Note: N/TERT-1 应在生物安全二级 (BSL-2) 条件下操作。该细胞系是研究银屑病、特应性皮炎等皮肤病病理机制的极佳体外工具。N/TERT-1 should be handled under Biosafety Level 2 (BSL-2) conditions. This cell line is an excellent in vitro tool for investigating the pathological mechanisms of skin diseases such as psoriasis and atopic dermatitis.

BioVector NTCC质粒载体菌株细胞蛋白抗体基因保藏中心

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