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A feasible method for the isolation of mesenchymal stem cells from menstrual blood and their exosomes
Razieh Dalirfardouei1, Khadijeh Jamialahmadi2, Elahe Mahdipour1
1Department of Medical Biotechnology, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran.
Background:
Mesenchymal stem cells (MSCs) are currently the most promising candidates in regenerative medicine. Nonetheless, there are several limitations associated with the MSC tissue source such as infrequent and invasive bone marrow sampling methods. To overcome these limitations, we have procured MSCs from the menstrual blood (MenSCs) as a non-invasive source using a straightforward and cost-effective method. Moreover, we isolated MenSCs-derived exosomes as a safe and highly effective approach to exert the paracrine effects of MSCs.
Methods:
MSCs were isolated from menstrual blood through two different culture methods: ficoll-isolated mononuclear cells (MNCs) and whole blood culture. These cells were characterized by their plastic adherence, flow cytometry analysis of the surface markers and the differentiation potential. The exosomes were isolated from conditioned media using ultracentrifugation and characterized by different microscopy techniques, western blotting, and ELISA.
Results:
Both Methods resulted in the rapid isolation of cells with MSC properties. However, the cellular yield of the whole blood culture method was remarkably more than MNCs culture. MenSCs also produced a substantial amount of extracellular vesicles (EVs) possessed the minimum criteria for exosome definition.
Conclusion:
The results suggest that direct culture of whole menstrual blood cells is a high throughput, straightforward and cost-effective method for MenSCs isolation using no special growth factors. Moreover, these cells are a good producer of exosome which can offer a cell-free therapy approach.
Insights
Menstrual blood stem cells (MenSCs) offer a non-invasive alternative to bone marrow stem cells for regenerative medicine. MenSCs are easily isolated from whole blood cultures and are potent producers of exosomes for cell-free therapy.
Area of Science:
- Regenerative Medicine
- Stem Cell Biology
- Exosome Biology
Background:
- Mesenchymal stem cells (MSCs) are key in regenerative medicine but face sourcing limitations due to invasive procedures.
- Menstrual blood stem cells (MenSCs) provide a non-invasive, cost-effective alternative MSC source.
- MenSCs-derived exosomes offer a safe and effective cell-free therapeutic approach.
Purpose of the Study:
- To establish a straightforward and cost-effective method for isolating MSCs from menstrual blood.
- To characterize MenSCs and their potential for exosome production.
- To evaluate MenSCs-derived exosomes for regenerative medicine applications.
Main Methods:
- MSCs were isolated from menstrual blood using two methods: ficoll-isolated mononuclear cells (MNCs) and whole blood culture.
- Cells were characterized by plastic adherence, surface marker flow cytometry, and differentiation potential.
- Exosomes were isolated via ultracentrifugation and characterized using microscopy, western blotting, and ELISA.
Main Results:
- Both isolation methods yielded cells with MSC properties.
- Whole blood culture produced a significantly higher cellular yield compared to MNC culture.
- MenSCs efficiently produced extracellular vesicles (EVs) meeting exosome criteria.
Conclusions:
- Direct culture of whole menstrual blood is a high-throughput, simple, and economical method for MenSCs isolation.
- MenSCs are excellent producers of exosomes, enabling a promising cell-free therapy strategy.
- This approach overcomes limitations of traditional MSC sourcing and offers a novel therapeutic avenue.
