Cell Line Development Knowledge Center

Reagents Help HEK Cell Line Viability And Viable Cell Density (VCD) During Cryopreservation And Revival

Cells often need a little extra support when it comes to stressful processes, such as cryopreservation and revival. Here we have a cell banking study which was conducted using ACF InstiTHAW HEK.

Cells were cryopreserved and revived with medium with and without InstiTHAW HEK to determine the supplements’ ability to support the viability and viable cell density (VCD) of cells.

Method

FreeStyle™ 293-F cells were cryopreserved at 1×106 cells/mL under the following conditions:

  • FreeStyle 293 expression medium supplemented with 10% DMSO, and InstiTHAW HEK.
  • FreeStyle 293 expression medium supplemented with 10% DMSO.

Following cryopreservation, the cells were revived under the following conditions:

  • FreeStyle 293 expression medium supplemented with InstiTHAW HEK.
  • FreeStyle 293 expression medium.

The viability (%) and VCD (cells/mL) of the cells cryopreserved and revived under the various conditions were assessed via Trypan Blue exclusion on day 6 post-revival.

Results

As you can see, the cells really benefitted from having InstiTHAW present during both cryopreservation and revival.

At day 6 post revival, the VCD of cells supplemented with InstiTHAW was 6.39×106 cells/mL compared to 3.55×106 cells/mL. Once more, the viability was also higher – Cells supplemented with InstiTHAW had a viability of 93% compared to 86% (see Figure 1 for more details).

Figure 1. Shows the viable cell density (cells/mL) and viability (%) of the FreeStyle™ 293-F cells day 6 post revival.

Thank you for joining us to review this case study. Please reach out to us at [email protected] if you would like to learn more about how InstiTHAW can help optimise our cell banking processes.

 

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