Improved Cryopreservation of Umbilical Cord Blood MSCs with Low‑dose DMSO

Vahidi, Mahmoud; Mozayyeni, Hossein; Mohajeri Iravani, Mojgan; Emamie, Amir; Ghorbani, Mahdi

doi:10.32598/JTRM.1.1006

Improved Cryopreservation of Umbilical Cord Blood MSCs with Low‑dose DMSO

Document Type : Original Article

Authors

Mahmoud Vahidi ¹

Hossein Mozayyeni ²

Mojgan Mohajeri Iravani ³

Amir Emamie ⁴

Mahdi Ghorbani ¹

¹ Cancer Epidemiology Research Center, AJA University of Medical Sciences, Tehran, Iran.

² Trauma and Surgery Research Center, AJA University of Medical Sciences, Tehran, Iran.

³ Department of Anesthesiology, Faculty of Paramedical, Hajar Hospital, AJA University of Medical Sciences, Tehran, Iran.

⁴ Department of Pathobiology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran.

https://doi.org/10.32598/JTRM.1.1006

Abstract

Background: Given the challenges in isolating mesenchymal stem cells (MSCs) from bone marrow (BM), umbilical cord blood (UCB) can be a promising source of MSCs. On the other hand, cryopreservation of adherent MSCs is a solution for long-term storage of these cells and subsequent experimental use of them. In the present study, we investigated the isolation and identification of UCB-MSCs and the effect of cell freezing with dimethyl sulfoxide (DMSO).
Methods: The successfully isolated UCB-MSCs were cultured in DMEM containing 15% fetal bovine serum and 1% penicillin-streptomycin. After flow cytometric analysis of the cells, we investigated the differentiation potential of MSCs. Finally, the CFU-F assay was performed before and after freezing with 5% and 10% DMSO.
Results: Results showed that the average number of mononuclear cells obtained from 12 UCB samples was 58.2±10.7×106 with a viability rate of 90±3%. MSCs were successfully isolated with a 33% recovery rate. These cells had fibroblastic-like morphology and were immunophenotypic, with adipogenic, osteogenic, chondrogenic, and neural differentiation capacities.
Conclusion: Based on the results, the use of 5% DMSO for UCB-MSC cryopreservation is recommended as an alternative to the conventional 10% DMSO. The UCB should be considered a promising alternative to BM as a source of MSCs. The use of the slow-freezing method with two concentrations of DMSO is effective in retaining the proliferation, cell-surface markers, and differentiation ability of human UCB-MSCs.

Keywords

Umbilical cord blood

Mesenchymal stem cells

Dimethyl sulfoxide (DMSO)

Differentiation

Freezing