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查看更多产品信息 Dynamis™ AGT™ Medium - FAQs (A2617503, A2617502, A2617504, A2617501)
4 个常见问题解答
请参考如下的逐步适应步骤：如果您在切换至某一比例时培养时遇到问题，请以前面的比例进行传代2-3次。
1.对培养在含5-10%血清的传统培养基或其他无血清培养基中的CHO细胞进行逐步适应培养时，请使用3×10E5–4×10E5 活性细胞/mL的接种密度。
2.使用Countess自动细胞计数仪监控细胞生长情况，直至活性细胞密度达到≥1×10E6个细胞/mL。
3.按照Dynamis AGT完全培养基：原始培养基的溶液25:75的比例来稀释细胞。我们推荐以原始比例的培养基培养一份细胞备份，直至新比例的培养基获得成功。在每一次后续传代时，使用比例逐渐增加的DynamisAGT 培养基溶液稀释细胞（Dynamis AGT完全培养基:原始培养基=25:75，50:50，75:25，90:10，最后是100%的Dynamis AGT培养基）。每一步骤都可能需要数次传代，以获得稳定的生长效果。
4.在100%的Dynamis AGT培养基中几次传代之后，活细胞计数在接种后3-4天内可达到至少2×10E6个细胞/mL，细胞存活率在85%以上。达到这一阶段，我们认为细胞完全适应了Dynamis AGT培养基。
We recommend using EfficientFeed A+, B+, and/or C+ AGT Supplements with Dynamis AGT Medium (Cat. Nos. A2617501, A2617502, A2617503, A2617504) to maximize titers. We recommend trying EfficientFeed C+ AGT Supplement (Cat. Nos. A2503101, A2503104, A2503105) and EffiientFeed C+ 2X Supplement (Cat. Nos. A3937601, A3937602) first, and if specific productivity drops off late in culture, try the addition of FunctionMAX TiterEnhancer (Cat. Nos. A1501001, A1501002). Supplements should not be added before half-peak cell density is reached for cultures not supplemented with complete feeding supplements like the EfficientFeed supplements.
Note: When culturing cells without complete feeds for fed-batch production we recommend that you not to allow glucose levels to fall below 2g/L in culture by supplementing with glucose as needed (see the product User Guide for more details).
Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.
When using Dynamis AGT Medium (Cat. Nos. A2617501, A2617502, A2617503, A2617504) for culturing a CHO cell line other than the glutamine synthetase (GS) gene expression system (GS-CHO expression system), the addition of 2-8 mM L-glutamine or a similar amount of GlutaMAX Supplement (Cat. No. 35050079, 35050061) is recommended.
Note: We recommend that you supplement with glucose as needed so that glucose levels don’t fall below 2g/L in culture (see the product User Guide for more details).
Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.
Please refer to the sequential adaptation procedure shown below. If you have trouble getting past a certain ratio, passage the cells 2-3 more times at the previous ratio.
1. During sequential adaptation of CHO cells grown in conventional 5-10% serum-supplemented medium or other serum-free medium, use a seeding density of 3 X 10E5-4 X 10E5 viable cells/mL.
2. Monitor cell growth using Countess Automated Cell Counter until viable cell density reaches greater than or equal to 1 X 10E6 cells/mL.
3. Dilute cells using a 25:75 ratio of complete Dynamis AGT Medium to the original medium. We recommend maintaining backup cultures in the original ratio medium until success with the new ratio medium is achieved. At each subsequent passage, dilute cells with stepwise increasing ratios of complete Dynamis AGT Medium to original medium (25:75, 50:50, 75:25, 90:10, followed by 100% Dynamis AGT Medium). Multiple passages at each step may be needed to achieve consistent growth.
4. After several passages in 100% Dynamis AGT Medium, the viable cell count should reach at least 2 X 10E6 cells/mL with ≥85% viability within 3-4 days of seeding culture. At this stage, the culture is considered to be adapted to Dynamis AGT Medium.
Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.