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Aama Javadi Rad1 
, Mohammad Khani-Eshratabdi2 , Morteza Zarrabi3 , Narjes Seddighi4 , Seyed Hadi Mousavi *5
, Mohammad Khani-Eshratabdi2 , Morteza Zarrabi3 , Narjes Seddighi4 , Seyed Hadi Mousavi *5
1- Department of Hematology and Blood Transfusion, School of Allied Medical Sciences, Tehran University of Medical Sciences, Tehran, Iran.
2- Department of Medical Laboratory Sciences, Kashmar School of Nursing, Mashhad University of Medical sciences (MUMS), Mashhad, Iran.
3- Department of Regenerative Medicine, Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran.
4- Student Research Committee, Tabriz University of Medical Sciences, Tabriz, Iran
5- 6. Department of Hematology and Blood Transfusion, School of Allied Medical Sciences, Tehran University of Medical Sciences, Tehran, Iran. ,hmousavi@tums.ac.ir
2- Department of Medical Laboratory Sciences, Kashmar School of Nursing, Mashhad University of Medical sciences (MUMS), Mashhad, Iran.
3- Department of Regenerative Medicine, Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran.
4- Student Research Committee, Tabriz University of Medical Sciences, Tabriz, Iran
5- 6. Department of Hematology and Blood Transfusion, School of Allied Medical Sciences, Tehran University of Medical Sciences, Tehran, Iran. ,
Abstract: (18 Views)
Background and objective: Exosomes derived from umbilical cord blood mesenchymal stem cells (UCB-MSCs) may enhance hematopoietic stem cells (HSCs) proliferation while preserving their undifferentiated state. This study evaluates the potential of UCB-MSC-derived exosomes as a cell-free strategy to expand HSCs, addressing the limitation of low HSC numbers for hematological disorder therapies.
Methods: In this in vitro study, MSC-derived exosomes (MSC-EXOs) were isolated and characterized. Subsequently, three umbilical cord blood samples were collected, and CD34+ HSCs were purified using magnetic-activated cell sorting (MACS). To identify the optimal exosome concentration, CD34+ cells were cultured with MSC-EXOs at four different concentrations (1, 5, 10, and 100 ng/mL). Gene expression levels of CXCR4, VLA-4, VLA-5, hTERT, and HOXA9 were assessed using quantitative RT-PCR, while colony-forming cell (CFC) assays were performed to evaluate the clonogenic potential of HSCs.
Results: After 7 days of co-culture, an increase in mononuclear cells (MNCs) was observed across all groups. Notably, the expression of the CD34 marker was significantly highest in Group III (71%) and Group IV (approximately 48%), exceeding levels seen in the other groups. Gene expression analysis revealed a significant upregulation of target genes in Group IV. Consistent with these findings, the CFC assay showed that Group IV had a significantly higher colonization rate than the other experimental conditions.
Conclusion: The results demonstrate that MSC-derived exosomes can effectively mimic the supportive functions of their parent cells in vitro, thereby significantly enhancing HSC proliferation.
Methods: In this in vitro study, MSC-derived exosomes (MSC-EXOs) were isolated and characterized. Subsequently, three umbilical cord blood samples were collected, and CD34+ HSCs were purified using magnetic-activated cell sorting (MACS). To identify the optimal exosome concentration, CD34+ cells were cultured with MSC-EXOs at four different concentrations (1, 5, 10, and 100 ng/mL). Gene expression levels of CXCR4, VLA-4, VLA-5, hTERT, and HOXA9 were assessed using quantitative RT-PCR, while colony-forming cell (CFC) assays were performed to evaluate the clonogenic potential of HSCs.
Results: After 7 days of co-culture, an increase in mononuclear cells (MNCs) was observed across all groups. Notably, the expression of the CD34 marker was significantly highest in Group III (71%) and Group IV (approximately 48%), exceeding levels seen in the other groups. Gene expression analysis revealed a significant upregulation of target genes in Group IV. Consistent with these findings, the CFC assay showed that Group IV had a significantly higher colonization rate than the other experimental conditions.
Conclusion: The results demonstrate that MSC-derived exosomes can effectively mimic the supportive functions of their parent cells in vitro, thereby significantly enhancing HSC proliferation.
Keywords: Differentiation, Extracellular vesicle, Hematopoietic stem cell, Mesenchymal stem cell, Proliferation
Type of Study: Original Research Article |
Subject:
Medical Biology
Received: 2025/07/3 | Accepted: 2026/07/1
Received: 2025/07/3 | Accepted: 2026/07/1
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