Dear all,
I need to culture whole blood in a 37°C incubator for 24h prior to RBC lysis and analysis.
If blood is sampled in Na-Heparin tubes, i think that it is required to add additional Na Heparin to the cell culture vessel to prevent clotting during the 24h culture.
Have someone already done that ? I read somewhere that 3 IU/mL Na Heparin is enough. Do you agree ?
Thank you.
Best regards.
Dear Benjamin Tschumi
If you have drawn the sample in the sodium-heparin tube then you need not to add further Na. Sodium in the tube is enough to stop coagulation.
Even if the coagulation will occur it will occur in 30-40 minutes. Check with control sample and then think of adding more Na to the tube.
Hope this is helpful.
Good luck.
Hi Benjamin Tschumi . You might be OK if you keep the blood in the collection tube.
I would not recommend putting whole blood on tissue culture plastic, if that's what you are thinking.
Steingrimur Stefansson Thank you for the insights.
Indeed, I planned to use plastic. Even with Heparin, it is not compatible ?
I will receive one tube of blood that I will split into different conditions.
What vessel do you recommend for culturing 24h at 37°C ?
Should I use an incubator with or without 5% CO2 ?
Benjamin Tschumi I would be concerned about platelet adhesion and activation on TC plastic.
I can't think of anything simple to prevent that, except fractionating the blood to get rid of most of the platelets before your experiments or using bacterial plate plastic, which does not support cell adhesion.
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