I'm trying to extract PBMC from New Zealand rabbit. I was wondering if anyone has some suggestions to increase the yield of PBMCs extraction from rabbit blood after Ficoll. I'm following the normal procedure but the final amount of cell is still highly contaminated with red blood cells and platelets. Usually the final amount of PBMCs contaminated with platelets is around 17-20 millions starting from 60ml of blood.
You can try treating your isolated PBMCs with Ammonium–chloride–potassium (ACK) as it helps breaks down the red blood cells. However, you will have to be careful while treating them as overtreatment can lead to a lower yield of your PBMCs. Here is a reference from my previous publication for this.
Ravishankar, P., Zeballos, M. A., & Balachandran, K. (2017). Isolation of endothelial progenitor cells from human umbilical cord blood. Journal of visualized experiments: JoVE, (127).
Alternatively, there are specialized centrifuge tubes available that would make the isolation easier as well. I have personally not used these but have heard from colleagues who have used them before. Here is a link to one of them, https://www.stemcell.com/products/brands/sepmate-pbmc-isolation.html
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