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Infusion of mature megakaryocytes into mice yields functional platelets
Rudy Fuentes, … , M. Anna Kowalska, Mortimer Poncz
Rudy Fuentes, … , M. Anna Kowalska, Mortimer Poncz
Published October 25, 2010
Citation Information: J Clin Invest. 2010;120(11):3917-3922. https://doi.org/10.1172/JCI43326.
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Brief Report

Infusion of mature megakaryocytes into mice yields functional platelets

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Abstract

Thrombopoiesis, the process by which circulating platelets arise from megakaryocytes, remains incompletely understood. Prior studies suggest that megakaryocytes shed platelets in the pulmonary vasculature. To better understand thrombopoiesis and to develop a potential platelet transfusion strategy that is not dependent upon donors, of which there remains a shortage, we examined whether megakaryocytes infused into mice shed platelets. Infused megakaryocytes led to clinically relevant increases in platelet numbers. The released platelets were normal in size, displayed appropriate surface markers, and had a near-normal circulating half-life. The functionality of the donor-derived platelets was also demonstrated in vivo. The infused megakaryocytes mostly localized to the pulmonary vasculature, where they appeared to shed platelets. These data suggest that it may be unnecessary to generate platelets from ex vivo grown megakaryocytes to achieve clinically relevant increases in platelet numbers.

Authors

Rudy Fuentes, Yuhuan Wang, Jessica Hirsch, Cheng Wang, Lubica Rauova, G. Scott Worthen, M. Anna Kowalska, Mortimer Poncz

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Figure 1

Characterization and infusion of megakaryocytes.

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Characterization and infusion of megakaryocytes.
(A) Representative fiel...
(A) Representative fields of small and large cells. Scale bars: 100 μm. (B) Representative analysis of DNA content of FL small and large cells. (C) Flow cytometry from recipient mouse before and after infusion of 108 WT platelets or (D) 106 FL large cells. (E) Flow cytometric percentage of 106 infused FL large cells and (F) 106 infused adult BM cells. n = 5 for WT platelets, n = 9 for FL cells, n = 5 for BM studies. (G) Percent platelet rise in irradiated thrombocytopenic mice after infusion. n = 5 per arm. Mean ± 1 SD are shown. Initial platelet counts (108/ml) in the 3 groups were: CATCH buffer, 1.8 ± 0.2; platelets, 1.9 ± 0.3; large cells, 1.0 ± 0.2. (H) Size determination of circulating recipient (blue) and infused platelets (red) by forward versus side scatter analysis. (I) Representative flow cytometric analysis of infused and FL-derived platelets comparing P-selectin, GPIbα, and GPIX.

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