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Retroviral Infection of Murine Embryonic Stem Cell Derived Embryoid Body Cells for Analysis of Hematopoietic Differentiation
Published on: October 20, 2014
Polycomb group protein Bmi1 promotes hematopoietic cell development from embryonic stem cells
Xiaolei Ding1, Qiong Lin, Roberto Ensenat-Waser
1Department of Cell Biology, Institute for Biomedical Engineering, RWTH Aachen University Medical School, Aachen, Germany.
Bmi1 (B-cell-specific Moloney murine leukemia virus integration site 1) enhances hematopoietic stem cell development from mouse embryonic stem cells. Forced Bmi1 expression promotes differentiation and expansion of hematopoietic progenitor cells.
Area of Science:
- Stem cell biology
- Epigenetics
- Hematopoiesis
Background:
- Bmi1 is a core component of Polycomb repressive complexes, crucial for adult stem cell maintenance and embryonic development.
- Polycomb repressive complexes regulate chromatin architecture and gene repression during development.
Purpose of the Study:
- To investigate the role of Bmi1 in mouse embryonic stem cells (ESCs) and during their differentiation.
- To determine the effect of forced Bmi1 expression on ESC self-renewal and hematopoietic differentiation.
Main Methods:
- Forced expression of Bmi1 in mouse ESCs.
- Global transcriptional profiling (RNA sequencing) to identify differentially regulated genes.
- Analysis of hematopoietic progenitor cell expansion and gene expression (Ink4a/Arf, Gata2).
Main Results:
- ESCs express low levels of Bmi1; forced expression does not affect self-renewal.
- Bmi1 enhances hematopoietic cell development upon ESC differentiation.
- Bmi1 up-regulates Gata2 expression, essential for primitive hematopoiesis.
- Bmi1 induces sustained growth and >100-fold expansion of ESC-derived hematopoietic stem/progenitor cells.
- Enhanced proliferation is linked to reduced Ink4a/Arf expression.
Conclusions:
- Bmi1 exhibits distinct roles in ESCs versus ESC-derived hematopoietic progenitor cells.
- Bmi1 promotes ESC differentiation towards the hematopoietic lineage.
- Bmi1 is a potential candidate gene for deriving adult stem cells from ESCs.
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