Hi all,
I am trying to reduce leukocyte contamination from my platelet-rich plasma sample from a dog.
If anyone has any experience in using mouse anti-CD45 microbeads for leukoreduction in a canine blood sample?
Thanks in advance.
Hi Antoni Iborra
,Data published by Karisch et al in 2011 suggests that CD45 may be expressed in platelets. Using mass spectrometry, they show the presence of a c-terminal peptide of CD45. Conversely, in 1988, Terstappen and Loken established that CD45 is not expressed on the surface of platelets by using flow cytometry.
But, very recently, Satya P. Kunapuli's lab demonstrated that platelets express a truncated CD45 isoform of approximately 65 kDa which has the catalytic cytoplasmic domain but lacks the extracellular domain and affects GPVI-mediated signaling in platelet.
Still, CD45 is currently being used for leukoreduction in PRP/PBMS samples in humans and mice, especially for proteomics studies to eliminate leukocyte contamination.
I wondered if the same product can be used for the canine samples as well!
The answer to the second question is:
Yes, we can do double centrifugation to reduce some amount of leukocyte contamination from the platelet. However, I am not sure of the total/99% of reduction, which may affect the data analysis further. Moreover, platelets are sensitive cells that get activated easily, and thus, I might not get my intended results from the experiment.
Hope this helps!