Abstract | BACKGROUND AIMS: Platelet transfusion can be a life-saving procedure in different medical settings. Thus, there is an increasing demand for platelets, of which shelf-life is only 5 days. The efficient ex vivo biomanufacturing of platelets would allow overcoming the shortages of donated platelets. METHODS: We exploited a two-stage culture protocol aiming to study the effect of different parameters on the megakaryo/thrombopoiesis ex vivo. In the expansion stage, human umbilical cord blood (UCB)-derived CD34(+)-enriched cells were expanded in co-culture with human bone marrow mesenchymal stromal cells (BM-MSCs). The megakaryocytic commitment and platelet generation were studied, considering the impact of exogenous addition of thrombopoietin (TPO) in the expansion stage and a cytokine cocktail (Cyt) including TPO and interleukin-3 in the differentiation stage, with the use of different culture medium formulations, and in the presence/absence of BM-MSCs (direct versus non-direct cell-cell contact). RESULTS: Our results suggest that an early megakaryocytic commitment, driven by TPO addition during the expansion stage, further enhanced megakaryopoiesis. Importantly, the results suggest that co-culture with BM-MSCs under serum-free conditions combined with Cyt addition, in the differentiation stage, significantly improved the efficiency yield of megakaryo/thrombopoiesis as well as increasing %CD41, %CD42b and polyploid content; in particular, direct contact of expanded cells with BM-MSCs, in the differentiation stage, enhanced the efficiency yield of megakaryo/thrombopoiesis, despite inhibiting their maturation. CONCLUSIONS: The present study established an in vitro model for the hematopoietic niche that combines different biological factors, namely, the presence of stromal/accessory cells and biochemical cues, which mimics the BM niche and enhances an efficient megakaryo/thrombopoiesis process ex vivo.
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Authors | Javad Hatami, Pedro Z Andrade, António Pedro Alves de Matos, Dusan Djokovic, Carla Lilaia, Frederico Castelo Ferreira, Joaquim M S Cabral, Cláudia L da Silva |
Journal | Cytotherapy
(Cytotherapy)
Vol. 17
Issue 4
Pg. 428-42
(Apr 2015)
ISSN: 1477-2566 [Electronic] England |
PMID | 25680300
(Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
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Copyright | Copyright © 2015 International Society for Cellular Therapy. Published by Elsevier Inc. All rights reserved. |
Chemical References |
- Antigens, CD34
- IL3 protein, human
- Interleukin-3
- Thrombopoietin
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Topics |
- Antigens, CD34
(metabolism)
- Blood Platelets
(cytology)
- Cell Communication
(physiology)
- Cell Differentiation
(drug effects)
- Cell Proliferation
(drug effects)
- Cells, Cultured
- Coculture Techniques
- Fetal Blood
(cytology)
- Hematopoietic Stem Cells
(cytology)
- Humans
- Interleukin-3
(pharmacology)
- Megakaryocytes
(cytology)
- Mesenchymal Stem Cells
(cytology)
- Platelet Transfusion
(methods)
- Thrombopoiesis
(physiology)
- Thrombopoietin
(pharmacology)
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