Harvesting HEK cells

Cell & Tissue Culture/ Cell Based Assays Cell Lines
1

Hello,

Currently I am working with HEK cells and since i have to study a cell surface receptor I am not using Trypsin EDTA for harvesting them. Instead i am using PBS-EDTA(0.02 %). The cells do dislodge after keeping the flask in the incubator for 1 min but I am unable to get a single cell suspension.

If anybody has faced similar problems or has a solution to this kindly reply soon.

Regards,

2

Leemark

Your options are limited since you are working on Cell Surface Receptor. However, you can still get this done effectively using a Trypsin free system.

  • Increase the incubation time and keep your cells at 37C. You may also increase the conc. of EDTA to .05M if that doesn’t interfere with your cells or downstream processing.
  • Triturate your cells using pipette gently which should dislodge the cellular clumps. Avoid vigorous pipetting as it may kill the cells
  • Use a sterile Sieve or mesh and pass your cells through it after dislodging from the cell culture flask.

Hope this will help you! Let us know in case you need further help.

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3

Try using an incubation with Accumax, sourced from Thermo Fisher.

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4

Nolan What are the constituents of Accumax?

5

The formulation is not listed on the website. But it’s fairly cheap and worth a try. Works great for accurately counting HEK cells. Accumax - Cell Aggregate Dissociation Medium

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