Article ; Online: Rapid isolation of mature murine primary megakaryocytes by size exclusion via filtration.
2023 Volume 34, Issue 1, Page(s) 2192289
Abstract: Megakaryocytes (MKs), the largest and rarest cells of the hematopoietic system, differentiate by increasing their size, DNA and cytoplasmic contents during maturation in order to release high numbers of blood platelets into the circulation. The gold- ... ...
| Abstract | Megakaryocytes (MKs), the largest and rarest cells of the hematopoietic system, differentiate by increasing their size, DNA and cytoplasmic contents during maturation in order to release high numbers of blood platelets into the circulation. The gold-standard to study these complex cells is the isolation of primary MKs from the native bone marrow (BM). This is typically achieved by using fluorescence- or magnetic-activated cell sorting. However, both methods are time-consuming and require a trained experimenter who is able to operate highly priced special equipment. Here, we demonstrate a simple and rapid alternative method to enrich mature MKs (≥16 N) from murine adult BM by size exclusion. The purity of the MK fraction reached 70-80% after isolation (100- to 250-fold enrichment). Reanalysis of isolated MKs by confocal microscopy revealed the expected expression of lineage-defining MK- and platelet-specific surface receptors, including CD42a/b/d and CD41/CD61. In addition, we detected a clear enrichment of MK-specific proteins/transcripts like |
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| MeSH term(s) | Humans ; Adult ; Animals ; Mice ; Megakaryocytes/metabolism ; Blood Platelets/metabolism ; Platelet Glycoprotein GPIb-IX Complex/metabolism |
| Chemical Substances | Platelet Glycoprotein GPIb-IX Complex |
| Language | English |
| Publishing date | 2023-03-30 |
| Publishing country | England |
| Document type | Journal Article |
| ZDB-ID | 1034283-7 |
| ISSN | 1369-1635 ; 0953-7104 |
| ISSN (online) | 1369-1635 |
| ISSN | 0953-7104 |
| DOI | 10.1080/09537104.2023.2192289 |
| Database | MEDical Literature Analysis and Retrieval System OnLINE |
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