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Measurement of Mitochondrial Mass and Membrane Potential in Hematopoietic Stem Cells and T-cells by Flow Cytometry
Published on: December 26, 2019
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Cells with hematopoietic potential reside within mouse proepicardium
Ewa Jankowska-Steifer1, Justyna Niderla-Bielińska2, Bogdan Ciszek3
1Department of Histology and Embryology, Center for Biostructure, Medical University of Warsaw, Chalubińskiego 5, 02-004, Warsaw, Poland.
Histochemistry and Cell Biology
|March 18, 2018
Summary
Mouse proepicardium contains endothelial cells with hematopoietic potential. These cells can differentiate into various blood cell types in vitro, suggesting a role in early hematopoiesis.
Area of Science:
- Developmental biology
- Hematopoiesis
- Endothelial cell differentiation
Background:
- Hematopoietic cells originate from hemogenic endothelium during embryonic development.
- The mouse proepicardium contains endothelial cells forming vascular networks.
- The hematopoietic potential of proepicardial endothelial cells is unexplored.
Purpose of the Study:
- To investigate the in vitro hematopoietic potential of proepicardial cell populations.
- To identify cell surface markers and gene expression patterns associated with hematopoietic differentiation from proepicardial cells.
Main Methods:
- Isolation and culture of proepicardial cell populations (CD31+/CD45-/CD71- and Flk-1+/CD31-/CD45-/CD71-).
- Colony-forming unit assays (CFU-GEMM, CFU-GM, CFU-E) to assess hematopoietic potential.
- Immunohistochemistry and RT-PCR to analyze cell phenotypes and gene expression (WT1, Runx1, Zeb1, Notch1, Gata2, Sox17).
Main Results:
- Proepicardial CD31+/CD45-/CD71- cells generated numerous CFU-GEMM, CFU-GM, and CFU-E colonies.
- Immunohistochemistry confirmed the presence of various hematopoietic lineages within these colonies.
- WT1 co-localized with Runx1 and Zeb1 in the proepicardium; Runx1 was expressed in CD31+ endothelial cells.
Conclusions:
- Endothelial cells from mouse proepicardium possess in vitro and in situ hematopoietic potential.
- These findings suggest a role for proepicardial cells in early hematopoiesis, akin to hemangioblast derivation.
- Expression of key regulatory genes (Runx1, Notch1, Gata2, Sox17) supports hemogenic endothelium specification.
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