Abstract
The in vitro production of blood platelets for transfusion purposes is an important goal in the context of a sustained demand for controlled products free of infectious, immune and inflammatory risks. The aim of this study was to characterize human platelets derived from CD34+ progenitors and to evaluate their hemostatic properties. These cultured platelets exhibited a typical discoid morphology despite an enlarged size and expressed normal levels of the major surface glycoproteins. They aggregated in response to ADP and a thrombin receptor agonist peptide (TRAP). After infusion into NSG mice, cultured and native platelets circulated with a similar 24 h half-life. Notably, the level of circulating cultured platelets remained constant during the first two hours following infusion. During this period of time their size decreased to reach normal values, probably due to their remodeling in the pulmonary circulation, as evidenced by the presence of numerous twisted platelet elements in the lungs. Finally, cultured platelets were capable of limiting blood loss in a bleeding assay performed in thrombocytopenic mice. In conclusion, we show here that cultured platelets derived from human CD34+ cells display the properties required for use in transfusion, opening the way to clinical trials.
Highlights
The in vitro production of blood platelets for transfusion purposes is an important goal in the context of a sustained demand for controlled products free of infectious, immune and inflammatory risks
The diameter of Cultured platelets was enlarged by 1.5 fold as compared to Native platelets (4.9 ± 0.13 vs 3.3 ± 0.1 μm, n = 70 and 95 respectively, n = 3) (Fig. 1A,B)
Cultured platelets displayed signs of immaturity illustrated by an increased density of multivesicular bodies (MVB), which represent granules in formation (0.6 ± 0.01 Cultured Platelets vs 0.06 ± 0.01 Native platlets, n = 95 and 50 respectively, ***p < 0.001), and an enriched endoplasmic reticulum
Summary
The in vitro production of blood platelets for transfusion purposes is an important goal in the context of a sustained demand for controlled products free of infectious, immune and inflammatory risks. The aim of this study was to characterize human platelets derived from CD34+ progenitors and to evaluate their hemostatic properties These cultured platelets exhibited a typical discoid morphology despite an enlarged size and expressed normal levels of the major surface glycoproteins. A reasonable application might be envisaged for well-identified needs, as in cases of transfusion failure due to platelet alloimmunization or the development of refractoriness[1,2] In this objective, efforts to improve different aspects of in vitro platelet production are well warranted. We explored their capacity to recirculate and to ensure a hemostatic protection equivalent to that of donor-derived platelets
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