Related Experiment Videos
Effect of recombinant human megakaryocyte growth and development factor coupled with polyethylene glycol on the
E Snyder1, P Perrotta, H Rinder
1Blood Bank and the Department of Laboratory Medicine, Yale University School of Medicine, Yale-New Haven Hospital, Connecticut 06504, USA.
Background:
Platelet production is regulated by a thrombopoietic growth factor (Mpl ligand). The receptor for this platelet growth factor (Mpl) is expressed on the platelet surface membrane. A recombinant thrombopoietic cytokine, recombinant human megakaryocyte growth and development factor coupled with polyethylene glycol (PEG-rHuMGDF), was added to apheresis platelets in vitro to determine whether Mpl ligand-receptor binding produced any beneficial or adverse effect on the development of the platelet storage lesion during 5 days of storage.
Study Design And Methods:
This study was designed as a dose-response protocol to determine the effects of adding increasing concentrations of PEG-rHuMGDF (0.0 [control], 2.5, 25, and 250 ng/mL) to apheresis platelets stored in two types of plastic storage containers. The increasing concentrations of PEG-rHuMGDF used simulated the theoretical peak plasma level attained in vivo, with an intravenous dose of 0, 0.1, 1.0 and 10 microg per kg of PEG-rHuMGDF. The platelets were stored with agitation at 20 to 24 degrees C for 5 days. A battery of in vitro assays was performed on storage Days 1 and 5, including pH, blood gases, platelet count, lactate dehydrogenase, mean platelet volume, glucose, lactate, osmotic recovery, morphology score, CD62P, and one-dimensional polyacrylamide gel electrophoresis analyses.
Results:
Analysis of results on both Day 1 and Day 5 showed no significant differences among any of the three PEG-rHuMGDF doses and the control group, for any in vitro assay. One-dimensional polyacrylamide gel electrophoresis showed no changes among the platelet protein patterns for the three PEG-rHuMGDF doses studied or the control. Storage-induced changes, however, did occur equally in all four groups of platelets over the 5 days of storage.
Conclusion:
The addition to stored apheresis platelets of up to 10 microg per kg of PEG-rHuMGDF (250 ng/mL), followed by 5 days of storage at standard conditions, does not appear to promote or retard development of the platelet storage lesion.
Insights
Adding recombinant human megakaryocyte growth and development factor (PEG-rHuMGDF) to stored apheresis platelets did not affect the platelet storage lesion. This study found no beneficial or adverse effects of PEG-rHuMGDF on platelet quality during storage.
Area of Science:
- Hematology
- Transfusion Medicine
- Biotechnology
Background:
- Platelet production is regulated by thrombopoietin (Mpl ligand).
- The Mpl receptor is present on platelet surfaces.
- Recombinant human megakaryocyte growth and development factor (PEG-rHuMGDF) was investigated for its effects on stored platelets.
Purpose of the Study:
- To determine if Mpl ligand-receptor binding influences the platelet storage lesion.
- To assess the impact of PEG-rHuMGDF on apheresis platelet quality during 5 days of storage.
Main Methods:
- A dose-response study using PEG-rHuMGDF concentrations (0-250 ng/mL).
- Apheresis platelets stored in plastic containers at 20-24°C for 5 days with agitation.
- In vitro assays performed on storage days 1 and 5, including pH, blood gases, platelet count, LDH, MPV, glucose, lactate, osmotic recovery, morphology, CD62P, and protein analysis.
Main Results:
- No significant differences were observed in any in vitro assay between PEG-rHuMGDF treated groups and the control.
- One-dimensional polyacrylamide gel electrophoresis showed no changes in platelet protein patterns.
- Storage-induced changes occurred similarly across all groups over 5 days.
Conclusions:
- PEG-rHuMGDF, at concentrations up to 250 ng/mL, did not promote or retard the development of the platelet storage lesion.
- The addition of PEG-rHuMGDF to stored apheresis platelets has no apparent effect on platelet quality during storage.