GC-1 spg

GC-1 spg 细胞系是通过转染 pSV3-neo 质粒实现永生化的，pSV3-neo 质粒含有 SV40 大 T 抗原的编码序列和新霉素抗性。这种基因修饰不仅提供了对某些抗生素的抗性，还通过改变细胞周期调节促进了细胞的持续生长，从而绕过了原代细胞典型的海弗里克极限。这种永生化过程可使细胞保持增殖能力，同时保留精原细胞的关键表型特征。

从表型上看，GC-1 spg 细胞系显示出介于 B 型精原细胞和初级精母细胞之间的过渡阶段的特征，使其成为研究精子发生早期阶段的一个特别相关的模型。这种细胞表达两种睾丸特异性同种蛋白：细胞色素 c 和乳酸脱氢酶 C4。这些标记对于研究精子发生过程中的细胞代谢和能量管理至关重要，反映了生殖细胞中活跃的独特代谢途径。这些特异性同种蛋白的表达强调了该细胞系在探索睾丸细胞功能和发育的生化和生理方面的实用性。

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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