The Phenotypic And Functional Properties Of Mouse Yolk-Sac-Derived Embryonic Macrophages
| Autor: | Jean-Leon Thomas, Tegy J. Vadakkan, Ross A. Poché, Mary E. Dickinson, Nejla Yosef, June-Hee Park |
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| Jazyk: | angličtina |
| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Population Cell Culture Techniques Biology Article 03 medical and health sciences Mice medicine Macrophage Animals Progenitor cell Yolk sac education Molecular Biology Myeloid Progenitor Cells Yolk Sac Tube formation Erythroid Precursor Cells education.field_of_study Microglia Macrophages Cell Differentiation Cell Biology Flow Cytometry Hematopoietic Stem Cells Embryonic stem cell Coculture Techniques Cell biology Endothelial stem cell 030104 developmental biology medicine.anatomical_structure Phenotype Developmental Biology |
| Popis: | Macrophages are well characterized as immune cells. However, in recent years, a multitude of non-immune functions have emerged many of which play essential roles in a variety of developmental processes (Wynn, Chawla and J. W. Pollard, 2013; DeFalco et al., 2014). In adult animals, macrophages are derived from circulating monocytes originating in the bone marrow, but much of the tissue-resident population arise from erythro-myeloid progenitors (EMPs) in the extra-embryonic yolk sac, appearing around the same time as primitive erythroblasts (Schulz et al., 2012; Kierdorf et al., 2013; McGrath, Frame and Palis, 2015; Gomez Perdiguero et al., 2015; Mass et al., 2016). Of particular interest to our group, macrophages have been shown to act as pro-angiogenic regulators during development (Owen and Mohamadzadeh, 2013; DeFalco et al., 2014; Hsu et al., 2015), but there is still much to learn about these early cells. The goal of the present study was to isolate and expand progenitors of yolk-sacderived Embryonic Macrophages (EMs) in vitro to generate a new platform for mechanistic studies of EM differentiation. To accomplish this goal, we isolated pure (>98%) EGFP(+) populations by flow cytometry from embryonic day 9.5 (E9.5) Csf1r-EGFP(+/tg) mice, then evaluated the angiogenic potential of EMs relative to Bone Marrow-Derived Macrophages (BMDMs). We found that EMs expressed more pro-angiogenic and less pro-inflammatory macrophage markers than BMDMs. EMs also promoted more endothelial cell (EC) cord formation in vitro, as compared to BMDMs in a manner that required direct cell-to-cell contact. Importantly, EMs preferentially matured into microglia when co-cultured with mouse Neural Stem/Progenitor Cells (NSPCs). In conclusion, we have established a protocol to isolate and propagate EMs in vitro, have further defined specialized properties of yolk-sac-derived macrophages, and have identified EM-EC and EM-NSPC interactions as key inducers of EC tube formation and microglial cell maturation, respectively. |
| Databáze: | OpenAIRE |
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