Packaging functionally important plasma proteins into the α-granules of human-induced pluripotent stem cell-derived megakaryocytes
| Autor: | Nanyan Zhang, Peter J. Newman |
|---|---|
| Rok vydání: | 2019 |
| Předmět: |
Induced Pluripotent Stem Cells
0206 medical engineering Immunocytochemistry Biomedical Engineering Medicine (miscellaneous) 02 engineering and technology Cytoplasmic Granules Article law.invention Biomaterials 03 medical and health sciences Von Willebrand factor Confocal microscopy law Humans Platelet Receptor Induced pluripotent stem cell 030304 developmental biology Megakaryopoiesis 0303 health sciences biology urogenital system Chemistry Blood Proteins 020601 biomedical engineering Blood proteins Cell biology biology.protein Megakaryocytes |
| Zdroj: | Journal of Tissue Engineering and Regenerative Medicine. 13:244-252 |
| ISSN: | 1932-6254 |
| DOI: | 10.1002/term.2785 |
| Popis: | The contents of platelet α-granules arrive via a number of pathways; some are synthesized by megakaryocytes (MKs), for example, von Willebrand factor (VWF), whereas others are endocytosed from plasma, for example, fibrinogen (Fgn) and factor V (FV). Currently, almost all in vitro-induced pluripotent stem cell (iPSC)-derived MKs are generated under serum-free conditions, and their α-granule cargoes lack components that would normally be taken up from plasma during the course of megakaryopoiesis. How this might affect the ability of in vitro-derived platelets to contribute fully to haemostasis is not known. The purpose of this investigation was to examine whether "feeding" human plasma to iPSC-derived MKs might result in loading their α-granules with physiologically important proteins. iPSCs were differentiated to CD41+ /CD42b+ MKs using a serum-free protocol. The resulting MKs were polyploid, expressed a number of platelet-specific surface receptors, and spread on Fgn or collagen-coated surfaces. Reverse transcription-polymerase chain reaction analysis detected mRNA transcripts for FV and VWF but not Fgn chains. Fluorescence immunocytochemistry and confocal microscopy confirmed constitutive VWF distribution in granule-like structures in MKs cultured under plasma-free conditions, and the granules became positive for Fgn upon incubation with human plasma. iPSC-derived MKs showed a low level of constitutive FV expression that increased dramatically upon incubation with human plasma. Taken together, these data suggest that human iPSC-derived MKs are capable of endocytosing and storing plasma components in their α-granules. Incorporating this methodology into current protocols for producing in vitro-derived MKs should provide novel insights into MK biology and lead to the generation of large numbers of MKs and platelets with improved functionality. |
| Databáze: | OpenAIRE |
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