Dear all,
I wonder whether you use antioxidants for cells before cryopreservation. Would you suggest a specific antioxidant? I found that (N-acetyl cysteine) was used for cryopreservation of human cord blood nucleated cells. I want to try it on PBMCs, do you have any ideas about the best concentration?
Thank you very much.
Hello Zain A. Karamya
I don’t use antioxidant for cryopreservation of human PBMCs. It is generally not required. I use a freezing medium containing 10% DMSO with 90% FBS which is a commonly used, inexpensive, and effective cryopreservation medium.
Application of antioxidants like N-acetyl cysteine is usually required when one has to maintain the survival and potency ability of cells (such as stem cells, human cord blood nucleated cells, etc.,) used for transplantation, especially when transplantation cannot be done immediately after cell harvest.
Antioxidant supplementation prior to cryopreservation is known to reduce oxidative stress and damage. Antioxidants protect cellular constituents from oxidative stress by neutralizing free radicals and terminating the oxidative chain reaction in mitochondrial membrane. N-acetyl cysteine is an acetylated cysteine residue containing sulfur, commonly used to inhibit ROS-induced cellular damage and apoptosis.
So, maintaining the quality and quantity of cells in terms of viability and functionality and making them available whenever required, the use of antioxidants becomes necessary. But still if you are interested to try it on the cryopreservation of human PBMCs, you may want to refer to the articles attached below. You could try 0.25uM - 0.5uM of N-acetyl cysteine.
https://www.ukm.my/jsm/pdf_files/SM-PDF-48-9-2019/15%20Shawal%20Maradona%20Abdul%20Wahab.pdf
Article The Role of N-Acetylcysteine Supplementation on the Oxidativ...
Best.
Malcolm Nobre I really appreciate your answer...thank you very much
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