Apheresis was carried out using a Spectra Optia v11.3 apheresis system (Terumo BCT, Inc Tokyo Japan) as detailed in the Additional file 11.

The continuous mononuclear cell collection procedure was used to sample from the targeted HCT layers from the blood of subjects 1 to 3. The targeted HCT layers that were sampled from these subjects ranged from the platelet rich layer through to the red cell rich layer.

A double stage procedure was used to collect the targeted HCT layers from subject 4. In the first stage, a red cell depletion procedure was used to collect approximately 500 ml of packed red blood cells from the subject. Targeted HCT layers were then collected from the red cell concentrate using the polymorphonuclear (PMN) collection procedure. The starting product (red cell concentrate) for the second stage of this procedure had a significantly higher HCT than the whole blood of subjects 1 to 3 and sampling focussed on the higher HCT layers. This was the rationale for using a PMN collection in subject 4 rather than the CMNC collection used in subjects 1 to 3.

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