Abstract
Mesenchymal stem cells (MSCs) are one of the most promising adult stem cells for clinical application in a cell therapy. The development of large-scale cryopreservation techniques, such as vitrification, for MSCs is a prerequisite for clinical therapies. Dimethyl sulfoxide (DMSO) and ethylene glycol (EG) are two types of cryoprotectants widely used for cell vitrification. However, the effects of DMSO and EG on the biological characteristics and transcriptome profiles of MSCs after cryopreservation remain unknown. In the present study, the viability, immunophenotype of cell surface markers, proliferation, differentiation potency, and global gene expression of rhesus macaque bone marrow-derived MSCs vitrified using DMSO and EG were studied. The results showed that vitrification did not affect the morphology, surface markers, and differentiation of the MSCs, and compared to DMSO, EG better protected cell viability and proliferation. Most importantly, vitrification resulted in changes in a large number of transcripts of MSCs either preserved using DMSO or EG. This report is the first to examine the effects of DMSO and EG on global gene expression in stem cells. These results will be beneficial to understanding the biological process involved in MSC vitrification and will contribute to improving cryopreservation protocols that maintain transcriptomic identity with high cryosurvival for preclinical research and clinical long-term storage.
Highlights
Mesenchymal stem cells are spindle-shaped fibroblast-like adult stem cells that are easy to isolate, culture, and expand in vitro
The surface marker profiles of the bone marrow-derived Mesenchymal stem cells (MSCs) were analyzed at passage 3 using flow cytometry
The MSCs derived from the bone marrow of macaques presented features of heterogeneous, fibroblastlike, and spindle-shaped morphology, consistent with the morphology of the bone marrow-derived MSCs as previously reported [33]
Summary
Mesenchymal stem cells are spindle-shaped fibroblast-like adult stem cells that are easy to isolate, culture, and expand in vitro. Mesenchymal stem cells can be differentiated into various cell types in vitro and in vivo under appropriate conditions, reflecting their multipotent capacity [1]. There are fewer ethical issues associated with MSCs than embryonic stem cells for clinical applications [2]. MSCs can be used in the treatment of a variety of clinical conditions and have been regarded as one of the most promising adult stem cells for clinical. Stem Cells International applications in cell therapy and regenerative medicine. The success of regenerative treatment with MSCs in clinical trials requires a large number of cells. The development of an optimal cryopreservation technique is a prerequisite for large-scale MSCs and storage for clinical therapies [6]
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