FreeStyle™ 293-F Cells - FAQs

View additional product information for FreeStyle™ 293-F Cells - FAQs (R79007)

10 product FAQs found

我没有FreeStyle 293-F细胞。我能在FreeStyle 293表达系统中使用一株其他的293细胞系吗?

其他的293细胞系也可能适用于FreeStyle 293表达系统。不过,它们需要先适应到FreeStyle 293表达培养基的无血清悬浮培养条件中,并需评估转染和表达效率。

从抗体和蛋白得率的角度来看,FreeStyle 293-F细胞与FreeStyle CHO-S细胞相比哪个更好?

当比较FreeStyle 293-F细胞与FreeStyle CHO-S细胞的平均抗体和蛋白得率时,绝对是FreeStyle 293-F细胞在瞬转条件下的抗体得率会比FreeStyle CHO-S细胞高。而另一方面,稳转CHO细胞的抗体得率会更高。对于非抗体蛋白而言,我们无法预测这两株细胞系的得率哪个更高。

培养于FreeStyle 293表达培养基中的FreeStyle 293-F细胞能使用ExpiFectamine293试剂和增强剂进行转染吗?如果可以,转染效率会比使用FreeStyle Max试剂更高吗?

您可使用ExpiFectamine293试剂来转染FreeStyle 293表达培养基中培养的FreeStyle 293-F细胞;不过,增强剂只会对蛋白表达提供少许的促进作用,因为这一产品是针对更高密度培养设计的,而FreeStyle 293表达培养基并不适用。 

I do not have FreeStyle 293-F cells. Can I use a different 293 cell line with the FreeStyle 293 Expression System?

Other 293 cell lines may be used with the FreeStyle 293 Expression System. However, they would have to be adapted to serum-free suspension culture in FreeStyle 293 Expression Medium and evaluated for transfection and expression.

How do FreeStyle 293-F cells compare to FreeStyle CHO-S cells in terms of antibody and protein yield?

When comparing FreeStyle 293-F cells with FreeStyle CHO-S cells for average range of antibody and protein yield, it is definitely expected that antibody yield will be higher in FreeStyle 293-F cells than in FreeStyle CHO-S cells, when generated by transient transfection. On the other hand, antibody yields in stable CHO cells can be much higher. For non-antibody proteins, it is impossible to predict which of the two cell lines may provide higher yields.

Can FreeStyle 293-F cells grown in FreeStyle 293 expression medium be transfected with ExpiFectamine293 Reagent along with enhancers? Is so, will the transfection efficiency be higher than that with FreeStyle Max Reagent?

You should be able to use the ExpiFectamine293 Reagent for transfection of Freestyle 293-F cells grown in FreeStyle 293 Expression Medium; however, the enhancers will provide only a little boost in expression as they are designed to work with higher density cultures that FreeStyle 293 Expression Medium cannot support.

Find additional tips, troubleshooting help, and resources within our Transfection Support Center.

How long can I keep culturing FreeStyle cells after transfection?

We typically keep the culture going for 7-10 days posttransfection. After that, protein expression may drop.

Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.

Is it necessary to grow FreeStyle 293-F cells in 8% CO2 when using the FreeStyle 293 Expression Medium? If 8% CO2 cannot be arranged, can I supplement the media with an additional reagent?

Our recommendation of 8% CO2 for the optimal growth of Freestyle 293 cells in Freestyle 293 Expression Medium is based on the bicarbonate content in the media and growth conditions of the cells observed by R&D. That said, you may use lower concentrations of CO2 if necessary (5% is common) to grow these cells. You may observe slightly slower growth kinetics or lower cell concentrations, but functionally the entire system will still work.

Why do FreeStyle 293-F cells require 8% CO2 for culturing?

The CO2 requirement is dependent on the media. Our serum-free 293 media require 8% CO2 to best buffer the media.

Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.

How are FreeStyle 293-F cells different from regular HEK293 cells?

FreeStyle 293-F cells are derived from HEK293 cells; they were just cloned extensively for growth, viability, and protein production in the new serum-free FreeStyle 293 medium. Thermo Fisher Scientific coined the name “FreeStyle” to signifiy the freedom of switching the cells to the defined serum-free FreeStyle media system.

The parental HEK293 cells, if adapted to the same media as FreeStyle 293F cells, should have the same transfection efficiency. However the final protein production levels may be very different as the FreeStyle 293F cells have been cloned and selected for maximal protein production.

Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.