Daoy

1985年，P.F. Jacobsen在西澳大利亚皇家珀斯医院建立了Daoy细胞系，这是一种从髓母细胞瘤中提取的人类细胞系，髓母细胞瘤是一种主要见于儿童的脑肿瘤。该细胞系源自一名 4 岁男孩的后窝肿瘤活检。髓母细胞瘤通常位于小脑，这是大脑中对运动控制和协调至关重要的区域，也是儿童最常见的恶性脑肿瘤。

Daoy细胞被广泛用作研究髓母细胞瘤生物学的模型系统，包括肿瘤的发生、发展和对疗法的反应。该细胞系在髓母细胞瘤研究中发挥了重要作用，尤其是在了解该疾病的分子和遗传基础以及测试化疗药物方面。这种细胞表现出恶性髓母细胞瘤的典型特征，包括生长速度快，移植到免疫力低下的小鼠体内能形成肿瘤。利用 Daoy 细胞系进行的研究有助于开发潜在的髓母细胞瘤新疗法和治疗靶点。

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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