Human platelet lysate is a rich source of growth factors and a potent supplement in cell culture. This protocol presents the process of preparing a large pool of human platelet lysate by starting from platelet rich plasma, performing several freeze-thaw cycles and depleting the platelet fragments.
In some regions platelet rich plasma (PRP) may be obtained from buffy-coats otherwise being a waste product of packed red blood cell production from tested blood donations (Figure 1). Optimally, PRP is used immediately for further preparation of pHPL, as in outdated platelet concentrates the availability of growth factors may be reduced due to platelet storage lesions and degradation20. It is further recommended to produce PRP by matching platelets of blood group O with plasma of blood group AB to avoid possib…
This work has been supported by the Austrian Research Foundation (FWF, grant N211-NAN to DS) and the Austrian Research Promotion Agency (FFG, grant N200 to DS). The authors thank Claudia Url for excellent technical assistance and Monica Farrell for linguistic editing.
Material Name | Tipo | Company | Catalogue Number | Comment |
---|---|---|---|---|
Double bag (2 x 3.5L) | MacoPharma | VDL 8000XQ | originally for ascites puncture | |
50 mL Falcon tube | Falcon BD | 2098 | ||
50 mL Stripettes | Costar | 4501 | ||
Plasma bags (600mL) | Baxter | R4R2021 | ||
Sterile tubing welder | Terumo | TSCD-II | ||
Welding equipment | Fresenius Kabi | Compo Seal | ||
Water bath | ||||
Sterile scissors | ||||
Clamps | ||||
Centrifuge |