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Co-culture of mesenchymal stem cell spheres with hematopoietic stem cells under hypoxia: a cost-effective method to maintain self-renewal and homing marker expression

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Abstract

Background

Hematopoietic stem cell (HSC) transplantation is considered a possible treatment option capable of curing various diseases. The aim of this study was the co-culturing of mesenchymal stem cell (MSC) spheres with HSCs under hypoxic condition to enhance the proliferation, self-renewal, stemness, and homing capacities of HSCs.

Methods and results

HSCs were expanded after being subjected to different conditions including cytokines without feeder (Cyto), co-culturing with adherent MSCs (MSC), co-culturing with adherent MSCs + hypoxia (MSC + Hyp), co-culturing with MSCs spheres (Sph-MSC), co-culturing with MSCs spheres + hypoxia (Sph-MSC + Hyp), co-culturing with MSC spheres + cytokines (Sph-MSC + Cyto). After 10 days, total nucleated cell (TNC) and CD34+/CD38− cell counts, colony-forming unit assay (CFU), long-term culture initiating cell (LTC-IC), the expression of endothelial protein C receptor (EPCR), nucleostemin (NS), nuclear factor I/X (Nfix) CXCR4, and VLA-4 were evaluated. The TNC, CD34+/CD38− cell count, CFU, and LTC-IC were higher in the Sph-MSC + Hyp and Sph-MSC + Cyto groups as compared with those of the MSC + Hyp group (P < 0.001). The expanded HSCs co-cultured with MSC spheres in combination with hypoxia expressed more EPCR, CXCR4, VLA-4, NS, and Nfix mRNA. The protein expression was also more up-regulated in the Sph-MSC + Cyto and Sph-MSC + Hyp groups.

Conclusion

Co-culturing HSCs with MSC spheres under hypoxic condition not only leads to higher cellular yield but also increases the expression of self-renewal and homing genes. Therefore, we suggest this approach as a simple and non-expensive strategy that might improve the transplantation efficiency of HSCs.

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Funding

This work was supported by the Hamadan University of Medical Sciences (Grant Number 990126370); and the Guilan University of Medical Sciences (Grant Number IR.GUMS.REC.1396.508).

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Authors and Affiliations

  1. Department of Medical Laboratory Sciences, School of Para Medicine, Hamadan University of Medical Sciences, Hamadan, Iran

    Fatemeh Amiri

  2. Department of Hematology and Blood Transfusion, Lorestan University of Medical Sciences, Khorramabad, Lorestan, Iran

    Ali Asghar Kiani

  3. Blood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine, Tehran, Iran

    Marzie Bahadori

  4. Cardiovascular Diseases Research Center, Department of Cardiology, Heshmat Hospital, School of Medicine, Guilan University of Medical Sciences, Rasht, Iran

    Mehryar Habibi Roudkenar

  5. Burn and Regenerative Research Center, Guilan University of Medical Sciences, Rasht, Iran

    Mehryar Habibi Roudkenar

Authors
  1. Fatemeh Amiri
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  2. Ali Asghar Kiani
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  3. Marzie Bahadori
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  4. Mehryar Habibi Roudkenar
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Contributions

Conceptualization: [FA and MHR], Methodology: [FA, AAK and MB], Formal analysis: [FA and MHR], Investigation and Writing—original draft preparation: [FA], Review and editing: [MB and MHR], Funding acquisition: [FA and MHR], Supervision: [MHR].

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Correspondence to Mehryar Habibi Roudkenar.

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The authors declare that they have no conflict of interest.

Ethical approval

All procedures were approved by Hamadan University of Medical Sciences Ethic Committee (No. IR.UMSHA.REC.1399.035). These procedures were in accordance with the ethical standards of the Institutional and/or National Research Committee and with the 1964 Helsinki Declaration and its later amendments or comparable ethical standards. Human umbilical cord tissue/blood was taken after filling of consent form.

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Amiri, F., Kiani, A.A., Bahadori, M. et al. Co-culture of mesenchymal stem cell spheres with hematopoietic stem cells under hypoxia: a cost-effective method to maintain self-renewal and homing marker expression. Mol Biol Rep 49, 931–941 (2022). https://doi.org/10.1007/s11033-021-06912-x

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