永生化 HK/FDC 细胞

永生化的HK/FDC细胞系是原始HK滤泡树突状细胞样细胞的遗传稳定衍生物，在保留关键表型和功能特征的同时，能够实现长期传代培养，且不受亲代培养物衰老相关的限制。通过引入特定遗传元件实现永生化，该方法可绕过复制阻滞，从而为长期研究生发中心生物学及FDC-B细胞相互作用提供了稳定平台。

永生化HK/FDC细胞在抗IgM或CD40配体等信号存在下，仍能结合并协同刺激生发中心B细胞，促进其存活并增强增殖能力。 关键的是，它们持续表达FDC特有的黏附分子和共刺激因子（包括VCAM-1和ICAM-1），并分泌可溶性介质，模拟原生FDC提供的微环境支持。这些特性使永生化HK/FDC细胞系成为解析B细胞在生发中心内成熟、亲和力选择及存活的细胞分子机制的可靠可重复模型。

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。

有关详细信息，请参阅我们的 <a href="" /Who-we-are/Warranty/"" hreflang=""x-default"">保修声明 。