Recent studies have identified a new human dental derived progenitor cell population with multi-lineage differentiation potential referred to as human periapical cyst mesenchymal stem cells (hPCy-MSCs). In the present study, we compared two subpopulations of hPCy-MSCs characterised by the low or high expression of CD146 to establish whether this expression can regulate their stem cell properties. Using flow cytometry, we evaluated the stem cell marker profile of hPCy-MSCs during passaging. Furthermore, CD146Low and CD146High cells were sorted by magnetic beads and subsequently both cell populations were evaluated for differences in their proliferation, self-renewal, stem cell surface markers, stemness genes expression and osteogenic differentiation potential.
We found that hPCy-MSCs possessed a stable expression of several mesenchymal stem cell surface markers, whereas CD146 expression declined during passaging.
In addition, sorted CD146Low cells proliferated significantly faster, displayed higher colony-forming unit-fibroblast capacity and showed higher expression of Klf4 when compared to the CD146High subset. Significantly, the osteogenic potential of hPCy-MSCs was greater in the CD146Low than in CD146High population. These results demonstrate that CD146 is spontaneously downregulated with passaging at both mRNA and protein levels and that the high expression of CD146 reduces the proliferative, self-renewal and osteogenic differentiation potential of hPCy-MSCs. In conclusion, our study demonstrates that changes in the expression of CD146 can influence the stem cell properties of hPCy-MSCs.
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The present study was supported by “ICARE Project - Infrastruttura Calabrese per la medicina Rigenerativa: “Generazione di biobanche per la criopreservazione di cellule staminali umane e di tessuto osseo per uso clinico e design e sviluppo di bioscaffold innovativi”. PON03PE_00009_2.2. The authors are grateful to Dr. Raghavendra Vasudeva Murthy and Dr. Pasquale Marrazzo for editing the manuscript.
Conflict of Interests
The authors declare that they have no conflicts of interest.
Francesco Paduano; Massimo Marrelli and Marco Tatullo equally contributed to this work.
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Paduano, F., Marrelli, M., Palmieri, F. et al. CD146 Expression Influences Periapical Cyst Mesenchymal Stem Cell Properties. Stem Cell Rev and Rep 12, 592–603 (2016). https://doi.org/10.1007/s12015-016-9674-4
- Dental stem cells (DSCs)
- Human periapical cyst mesenchymal stem cells (hPCy-MSCs)
- MSC surface markers
- Magnetic cell sorting
- Stemness genes
- Osteogenic differentiation