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Identification of PDGF receptors on human megakaryocytes and megakaryocytic cell lines
M Yang1, L M Khachigian, C Hicks
1The Centre for Thrombosis and Vascular Research, School of Pathology, The University of New South Wales and Department of Haematology, The Prince of Wales Hospital, Sydney, Australia.
Abstract:
Platelet-derived growth factor (PDGF) is a potent chemotactic and mitogenic factor implicated to play important roles in a variety of normal and pathophysiologic settings. We investigated PDGF receptor expression on human megakaryocytes and several megakaryocytic cell lines (CHRF, DAMI, Meg-01, M-07e) using enzyme-linked immunosorbent assay (ELISA), flow cytometry and immunocytochemical staining. Both PDGF receptor subtypes were identified on CHRF, DAMI, and Meg-01 cells by ELISA; PDGF beta-receptor levels exceeded alpha-receptor levels. Flow cytometry revealed that beta-receptor levels on CHRF and DAMI cells exceeded those on Meg-01 cells, and that M-07e expressed neither receptor. Immunocytochemical staining confirmed these findings and determined that bone marrow megakaryocytes also expressed PDGF receptors. Exposure of megakaryocytes to PDGF-BB dramatically induced the expression of the immediate-early gene, c-fos, within 30 min. Moreover, PDGF-BB significantly stimulated CHRF proliferation and colony formation. The present study demonstrates the presence of functional PDGF receptors on human megakaryocytes and their ability to mediate a mitogenic response.
Insights
Platelet-derived growth factor (PDGF) receptors are present on human megakaryocytes, influencing cell growth. This study confirms functional PDGF receptors mediate a mitogenic response in these critical blood cells.
Area of Science:
- Hematology
- Cell Biology
- Molecular Biology
Background:
- Platelet-derived growth factor (PDGF) is a key factor in cellular processes.
- Its role in megakaryocytes, the cells that produce platelets, is not fully understood.
Purpose of the Study:
- To investigate Platelet-derived growth factor (PDGF) receptor expression on human megakaryocytes.
- To determine if PDGF can stimulate megakaryocyte proliferation and gene expression.
Main Methods:
- Enzyme-linked immunosorbent assay (ELISA)
- Flow cytometry
- Immunocytochemical staining
- Gene expression analysis (c-fos)
- Cell proliferation and colony formation assays
Main Results:
- PDGF alpha- and beta-receptors were detected on human megakaryocyte cell lines (CHRF, DAMI, Meg-01), with higher beta-receptor levels.
- Bone marrow megakaryocytes also expressed PDGF receptors.
- PDGF-BB induced c-fos gene expression and significantly stimulated proliferation and colony formation in CHRF cells.
Conclusions:
- Human megakaryocytes express functional PDGF receptors.
- PDGF signaling mediates a mitogenic response in megakaryocytes, impacting platelet production.