NCI-H1781

NCI-H1781 细胞系是一种源自肺腺癌的人类非小细胞肺癌（NSCLC）模型。该细胞系尤其值得注意的是，它携带 ERBB2（HER2）G776insV_G/C 突变，这是 20 号外显子的框架内插入突变，具有激活功能。这种突变是已知的肺癌亚群的驱动因素，使 NCI-H1781 成为研究 HER2 靶向疗法和耐药机制的有用模型。NCI-H1781中的ERBB2突变有助于构成性激酶激活，并通过PI3K/AKT和MAPK等通路发出下游信号，从而支持细胞增殖和存活，而不受外部生长因子的影响。

在分子谱分析研究中，NCI-H1781 的 ERBB2 转录本和蛋白水平升高，这与其基因改变相一致。此外，这种细胞系经常被用于药物基因组学研究，因为它对拉帕替尼或阿法替尼等 HER2 抑制剂的敏感性会因细胞环境和组合靶向策略的不同而变化。它还表现出对表皮生长因子受体抑制剂的耐药性，这使它有别于表皮生长因子受体突变的肺癌模型，并强调了 HER2 特异性靶向的治疗意义。NCI-H1781 具有良好的遗传背景和体外生长特性，是测试 HER2 靶向化合物和探索肺腺癌耐药机制的可靠临床前模型。

37°C, 5%_CO2, 加湿环境。

1. Confirm that the vial remains deeply frozen upon delivery, as cells are shipped on dry ice to maintain optimal temperatures during transit.

2. Upon receipt, either store the cryovial immediately at temperatures below -150°C to ensure the preservation of cellular integrity, or proceed to step 3 if immediate culturing is required.

3. For immediate culturing, swiftly thaw the vial by immersing it in a 37°C water bath with clean water and an antimicrobial agent, agitating gently for 40-60 seconds until a small ice clump remains.

4. Perform all subsequent steps under sterile conditions in a flow hood, disinfecting the cryovial with 70% ethanol before opening.

5. Carefully open the disinfected vial and transfer the cell suspension into a 15 ml centrifuge tube containing 8 ml of room-temperature culture medium, mixing gently.

6. Centrifuge the mixture at 300 x g for 3 minutes to separate the cells and carefully discard the supernatant containing residual freezing medium.

7. Gently resuspend the cell pellet in 10 ml of fresh culture medium. For adherent cells, divide the suspension between two T25 culture flasks; for suspension cultures, transfer all the medium into one T25 flask to promote effective cell interaction and growth.

8. Adhere to established subculture protocols for continued growth and maintenance of the cell line, ensuring reliable experimental outcomes.

使用基于 PCR 的检测方法和基于发光的支原体检测方法排除支原体污染。

为确保没有细菌、真菌或酵母菌污染，每天都要对细胞培养物进行目视检查。

如需长期保存，可将样品瓶放入气相液氮中，温度约为 -150 至 -196 ℃。在-80 °C下保存只能作为转入液氮前的短暂过渡。

冷冻保存的细胞系用干冰装运，并用经过验证的隔热包装和足够的制冷剂在整个运输过程中保持约 -78 °C 的温度。收到货后，应立即检查容器，并立即将小瓶转移到适当的储藏室。

在Cytion，我们保证产品的质量和活力。 我们保证，从我们的工厂发货后 60 天内，我们的细胞在开始培养时都是存活的。 我们保证所有材料均符合分析证书和/或产品说明书中描述的规格。

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