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Updated: Feb 15, 2026
Preparation and Gene Modification of Nonhuman Primate Hematopoietic Stem and Progenitor Cells
Published on: February 15, 2019
Hematopoietic stem/progenitor cell senescence is associated with altered expression profiles of cellular
Yongpin Dong1, Xiaolan Lian2, Yanwu Xu3
1Department of Emergency and Critical Care Medicine, Shanghai Changzheng Hospital, The Second Military Medical University, Shanghai 200003, China.
Insights
Cellular memory mechanisms contribute to hematopoietic stem/progenitor cell (HSPC) senescence in aging mice. Altered gene expression in senescent HSPCs may impact stem cell plasticity and disease progression.
Area of Science:
- Hematology
- Cellular Biology
- Aging Research
Background:
- Hematopoietic stem/progenitor cells (HSPCs) undergo senescence with age.
- Cellular memory mechanisms are implicated in aging and stem cell function.
- Understanding HSPC senescence is crucial for addressing age-related diseases.
Purpose of the Study:
- To investigate the role of cellular memory mechanisms in HSPC senescence.
- To compare senescent markers and gene expression in young versus aged mouse HSPCs.
Main Methods:
- Purification of HSPCs (Lin-CD117+) from young and aged mice using Magnetic Activated Cell Sorting (MACS).
- Assessment of cell cycle distribution via flow cytometry.
- Evaluation of proliferative capacity using the CFU-Mix assay.
- Quantitative real-time PCR to measure mRNA levels of Polycomb Group (PcG) and Trithorax Group (TrxG) genes.
Main Results:
- Aged mice exhibited a higher percentage of senescent HSPCs, increased cell cycle arrest (G0/G1 phase), and reduced proliferation (CFU-Mix).
- Key cellular memory genes (Ezh1, Bmi-1, Eed, Rae-28) showed significantly lower mRNA expression in aged HSPCs.
- Mel18 mRNA expression was significantly higher in senescent HSPCs from older mice.
Conclusions:
- Altered expression of cellular memory-associated genes in senescent HSPCs suggests a role in aging.
- These molecular changes may impair the plasticity of aged hematopoietic stem cells.
- The findings contribute to understanding senescence-associated disease processes in the hematopoietic system.
Abstract:
To evaluate the contributions of cellular memory mechanisms to hematopoietic stem/progenitor cell (HSPC) senescence. HSPCs (Lin-CD117+, hereafter referred to as HSPC) were separated from young (6-week-old) and aged (18-month-old) mice using Magnetic Activated Cell Sorting (MACS). Cell cycle distribution of HSPCs was determined using flow cytometry. The mixed colony forming unit (CFU-Mix) assay was used to study the HSPCs' ability to proliferate. The mRNA expression levels of cellular memory-implicated PCG family (enhancer of zeste homolog 2 (Ezh2), B lymphoma mo-MLV insertion region 1 (Bmi-1), embryonic ectoderm development (Eed), melanoma nuclear protein 18 (Mel18), Mph1/polyhomeotic-like protein 1 (Rae-28)) and Trithorax group (TrxG) family (mixed lineage leukemia (Mll), thioredoxin (Trx)) were determined by quantitative real-time PCR. We obtained highly purified populations of mouse HSPCs (Lin-CD117+) (92.2 ± 4.5% CD117+). The percentage of HSPCs was significantly higher in older mice compared with younger control mice and the percentage of SA-β-galactosidase positive cells was significantly higher in HSPCs isolated from older mice (P<0.05). The percentage of HSPCs in G0/G1 was significantly higher in older mice compared with younger control mice (52.0 compared with 47.1%), indicating increased cell cycle arrest in senescent HSPCs. The amount of CFU-Mix was significantly decreased in aged group (13.8 compared with 40.0), indicating a diminished ability to proliferate in senescent HSPCs. Ezh1, Bmi-1, Eed, Rae-28 gene mRNA expression was significantly lower in HSPCs from older mice compared to younger controls, while Mel18 mRNA expression was significantly higher in HSPCs from older mice (P<0.05). The expression of genes associated with cellular memory is altered in senescent (Lin- CD117+) HSPCs, which may affect the potential plasticity of aged hematopoietic stem cells (HSCs) and thereby contribute to senescence-associated disease processes.
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