BioVector? hERG-HEK293 穩(wěn)定表達(dá)細(xì)胞系 / BioVector? hERG-HEK293 Stable Expression Cell Line

通用定義 / General Definition:

BioVector? hERG-HEK293 是一種穩(wěn)定轉(zhuǎn)染了人 hERG (human Ether-à-go-go-Related Gene) 鉀離子通道基因的 HEK293（人胚胎腎細(xì)胞） 衍生細(xì)胞系。hERG 通道負(fù)責(zé)心肌動(dòng)作電位復(fù)極化的關(guān)鍵電流（$I_{Kr}$）。該細(xì)胞系是制藥行業(yè)進(jìn)行**藥物安全性評(píng)價(jià)（心臟毒性篩查）**的“金標(biāo)準(zhǔn)”工具。幾乎所有新藥在臨床前階段都必須通過 hERG 極化實(shí)驗(yàn)，以評(píng)估其是否會(huì)引起 QT 間期延長(zhǎng)及潛在的致命性心律失常（如尖端扭轉(zhuǎn)型室性心動(dòng)過速）。

BioVector? hERG-HEK293 is a HEK293 (Human Embryonic Kidney) derivative cell line stably transfected with the human hERG (human Ether-à-go-go-Related Gene) potassium channel. The hERG channel is responsible for the rapid delayed rectifier potassium current ($I_{Kr}$) essential for cardiac action potential repolarization. This cell line is the "gold standard" tool in the pharmaceutical industry for drug safety assessment (cardiotoxicity screening). Nearly all drug candidates must undergo hERG testing during preclinical stages to evaluate their potential to cause QT interval prolongation and life-threatening arrhythmias, such as Torsade de Pointes.

BioVector? hERG-HEK293 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? hERG-HEK293 穩(wěn)定表達(dá)細(xì)胞系

• 母本細(xì)胞： HEK293

• 靶點(diǎn)基因： 人 KCNH2 (hERG)

• 生長(zhǎng)特性： 貼壁生長(zhǎng)

• 細(xì)胞形態(tài)： 上皮樣，呈多角形

2. 培養(yǎng)條件

• 基礎(chǔ)培養(yǎng)基： BioVector? DMEM 培養(yǎng)基（高糖）。

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

• 篩選抗生素： 含有適量的 G418 或 Hygromycin B（取決于載體構(gòu)建），以維持 hERG 通道的穩(wěn)定表達(dá)。

• 培養(yǎng)環(huán)境： 37 攝氏度，5% $CO_2$

• 傳代比例： 1:3 至 1:6；電生理實(shí)驗(yàn)建議在細(xì)胞處于對(duì)數(shù)生長(zhǎng)期時(shí)進(jìn)行。

3. 細(xì)胞應(yīng)用

• 電生理篩查： 利用手動(dòng)或自動(dòng)膜片鉗（Patch Clamp）技術(shù)檢測(cè)候選藥物對(duì) hERG 電流的抑制作用（$IC_{50}$ 值）。

• 心臟安全性評(píng)價(jià)： 遵循 GLP 規(guī)范，評(píng)估非心臟病藥物的潛在致心律失常風(fēng)險(xiǎn)。

• 通道功能研究： 研究 hERG 通道的門控機(jī)制、動(dòng)力學(xué)及輔助亞基的影響。

4. 注意事項(xiàng)

• 電流強(qiáng)度監(jiān)測(cè)： 隨代數(shù)增加，通道表達(dá)水平可能下降。BioVector? 建議定期通過膜片鉗檢測(cè)全細(xì)胞電流密度（通常要求應(yīng)大于 500 pA）。

• 消化處理： 膜片鉗實(shí)驗(yàn)需使用溫和的消化液（如低濃度 Trypsin 或 Accutase），以防止對(duì)膜蛋白受體的機(jī)械性損傷。

English Datasheet

1. General Product Information

• Product Name: BioVector? hERG-HEK293 Stable Expression Cell Line

• Parental Line: HEK293

• Target Gene: Human KCNH2 (hERG)

• Growth Properties: Adherent

• Morphology: Epithelial-like, polygonal.

2. Culture Conditions

• Basal Medium: BioVector? DMEM (High Glucose).

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

• Selection Marker: Maintain with G418 or Hygromycin B (dependent on the specific construct) to ensure stable hERG channel expression.

• Incubation: 37 degrees Celsius, 5% $CO_2$.

• Subculturing: 1:3 to 1:6 ratio; cells should be in the logarithmic growth phase for electrophysiological recording.

3. Applications

• Electrophysiological Screening: Utilizing manual or automated Patch Clamp techniques to determine $IC_{50}$ values for drug-induced inhibition of hERG currents.

• Cardiac Safety Pharmacology: Assessing pro-arrhythmic risks of non-cardiovascular drugs in compliance with GLP guidelines (e.g., ICH S7B).

• Channel Biophysics: Studying gating mechanisms, kinetics, and the influence of auxiliary subunits on hERG function.

4. Key Usage Notes

• Current Density Monitoring: Channel expression may diminish over long-term culture. BioVector? recommends periodic verification of whole-cell current amplitude (typically $> 500$ pA is required for high-quality data).

• Gentle Dissociation: For patch clamp assays, use mild dissociation reagents (e.g., Accutase) to preserve membrane integrity and channel protein functionality.

注意 / Note: BioVector? hERG-HEK293 細(xì)胞在傳代至 20 代后，電生理性能可能發(fā)生漂移。為確保數(shù)據(jù)精確度，建議及時(shí)從冷凍管復(fù)蘇低代數(shù)細(xì)胞。

The electrophysiological performance of BioVector? hERG-HEK293 may drift after 20 passages. To ensure data accuracy, BioVector? recommends thawing a new vial of low-passage cells periodically.

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