BioVector? NK3.3 人源自然殺傷細(xì)胞系 / BioVector? NK3.3 Human Natural Killer Cell Line

通用定義 / General Definition:BioVector? NK3.3 是一種具有高度細(xì)胞毒性的人源非轉(zhuǎn)化自然殺傷 (NK) 細(xì)胞系。該細(xì)胞系最初由 Jacki Kornbluth 實(shí)驗(yàn)室從一名健康志愿者的外周血中分離并建立。與常見的 NK 樣腫瘤細(xì)胞系（如 NK-92）不同，NK3.3 被認(rèn)為是非惡性轉(zhuǎn)化的，且在生長(zhǎng)過程中嚴(yán)格依賴白細(xì)胞介素-2 (IL-2)。NK3.3 保留了原代 NK 細(xì)胞的許多關(guān)鍵特征，包括對(duì)多種腫瘤靶細(xì)胞的自發(fā)殺傷活性，以及介導(dǎo)抗體依賴性細(xì)胞毒性 (ADCC) 的能力。它是研究 NK 細(xì)胞激活、抑制受體信號(hào)傳導(dǎo)以及開發(fā)新型免疫療法的極佳體外模型。

BioVector? NK3.3 is a highly cytotoxic, non-transformed human Natural Killer (NK) cell line. Originally isolated from the peripheral blood of a healthy donor by Jacki Kornbluth's laboratory, NK3.3 is distinct from common malignant NK cell lines (like NK-92) as it is considered non-transformed and exhibits a strict dependency on Interleukin-2 (IL-2) for growth. NK3.3 retains the key functional hallmarks of primary NK cells, including potent spontaneous cytotoxicity against various tumor targets and the ability to mediate Antibody-Dependent Cellular Cytotoxicity (ADCC). It serves as an excellent in vitro model for studying NK cell activation, inhibitory receptor signaling, and the development of novel immunotherapies.

BioVector? NK3.3 技術(shù)說明書 (Technical Datasheet)

中文版說明書 (Chinese Datasheet)

1. 產(chǎn)品基本信息

• 產(chǎn)品名稱： BioVector? NK3.3 人自然殺傷細(xì)胞

• 組織來源： 人外周血 (Peripheral Blood)

• 細(xì)胞特性： 非轉(zhuǎn)化、IL-2 依賴性、高度細(xì)胞毒性

• 表面標(biāo)志物： CD56+, CD16+, CD3- (典型的 NK 細(xì)胞表型)

• 主要用途： 細(xì)胞毒性實(shí)驗(yàn)、ADCC 研究、NK 細(xì)胞受體生物學(xué)

2. 培養(yǎng)條件

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

• 血清添加： 10%-15% BioVector? 優(yōu)質(zhì)胎牛血清 (FBS)。

• 關(guān)鍵添加劑： 必須添加重組人 IL-2 (通常為 100-200 U/mL)。

• 補(bǔ)充因子： 2 mM L-谷氨酰胺、1% 非必需氨基酸、50 uM beta-巰基乙醇。

• 培養(yǎng)環(huán)境： 37 攝氏度，5% CO2，懸浮培養(yǎng)。


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3. 細(xì)胞應(yīng)用

• 殺傷效率評(píng)價(jià)： 用于測(cè)試 NK 細(xì)胞對(duì) K562（對(duì) NK 敏感）等靶細(xì)胞的裂解能力。

• ADCC 實(shí)驗(yàn)： 評(píng)估治療性抗體（如利妥昔單抗）結(jié)合靶細(xì)胞后誘導(dǎo) NK3.3 殺傷的效果。

• 細(xì)胞因子產(chǎn)生： 刺激后產(chǎn)生 IFN-gamma 和 TNF-alpha 的能力。

4. 注意事項(xiàng)

• IL-2 敏感性： 缺乏 IL-2 會(huì)迅速導(dǎo)致細(xì)胞凋亡。建議每 2-3 天更換含新鮮 IL-2 的培養(yǎng)基。

• 細(xì)胞密度： 維持在 200,000 到 800,000 cells/mL 之間；密度過低會(huì)影響增殖，密度過高會(huì)導(dǎo)致活力下降。

English Datasheet

1. General Product Information

• Product Name: BioVector? NK3.3 Human Natural Killer Cell Line

• Source: Human peripheral blood

• Characteristics: Non-transformed, strictly IL-2 dependent, potent effector functions.

• Phenotype: CD56 positive, CD16 positive, CD3 negative.

• Primary Use: Assessing innate cytotoxicity, ADCC, and NK receptor signaling cascades.

2. Culture Conditions

• Basal Medium: BioVector? RPMI 1640.

• Serum Supplement: 10%-15% BioVector? Fetal Bovine Serum (FBS).

• Critical Factor: Supplemented with recombinant human IL-2 (100-200 U/mL).

• Supplements: 2 mM L-Glutamine, 1 mM Sodium Pyruvate, and 50 uM beta-mercaptoethanol.

• Incubation: 37°C, 5% CO2, suspension culture.


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3. Applications

• Cytotoxicity Assays: Standard effector for chromium release or flow-based killing assays against K562 or Raji cells.

• Immunotherapy Screening: Testing the efficacy of Bispecific Killer Cell Engagers (BiKEs) or TriKEs.

• Signal Transduction: Studying the phosphorylation of Syk, Zap70, and PLC-gamma upon activating receptor ligation.

4. Key Usage Notes

• Growth Kinetics: NK3.3 cells grow more slowly compared to malignant lines like NK-92. Patience is required during the initial expansion phase after thawing.

• Maintenance: Regularly monitor CD16 (Fc-gamma RIII) expression via flow cytometry if the cells are being used for ADCC assays, as expression levels can fluctuate.

注意 / Note: NK3.3 應(yīng)在生物安全二級(jí) (BSL-2) 條件下操作。該細(xì)胞系對(duì)操作過程中的物理壓力較為敏感，復(fù)蘇后建議通過緩慢添加培養(yǎng)基的方式進(jìn)行過渡。NK3.3 should be handled under Biosafety Level 2 (BSL-2) conditions. This cell line is sensitive to physical stress; BioVector? recommends a gradual medium transition and gentle handling during the post-thaw recovery period.

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