Isolation and identification of CXCR4-positive cells from human dental pulp cells
Long Jiang1, Wei-Wei Peng, Li-Fen Li
1Department of General Dentistry, 9th People's Hospital, Shanghai JiaoTong University, School of Medicine, Shanghai Key Laboratory of Stomatology, Shanghai, PR China.
Introduction:
In previous studies, we found expression of stromal cell-derived factor-1α (SDF-1α)/CXC chemokine receptor 4 (CXCR4) in human dental pulp and the SDF-1α-CXCR4 axis might play a role in the recruitment of CXCR4-positive dental pulp cells (CXCR4(+) DPCs) toward the damaged sites. However, the specific function of CXCR4(+) DPCs in the injured dental pulp was still unknown. The purpose of this study was to isolate CXCR4(+) DPCs from dental pulp cells in vitro to pave the way for further study of their characteristics.
Methods:
CXCR4(+) DPCs were isolated with magnetic-activated cell sorting (MACS). Freshly isolated CXCR4(+) DPCs were identified by immunohistochemistry with light microscopy or confocal microscopy. Then the phenotypes CXCR4, stromal cell surface marker-1 (STRO-1), CD146, and CD34 in 3 groups (ie, CXCR4(+) DPCs, CXCR4(-) DPCs, or non-sorted DPCs) were analyzed by flow cytometry after they were cultured and expanded in vitro.
Results:
The results indicated the isolated subpopulation of DPCs was enriched with CXCR4(+) DPCs, and the positive rates of STRO-1 and CD146 in CXCR4(+) DPCs group were higher than CXCR4(-) DPCs or non-sorted DPCs groups (P < .05). There was no expression of CD34 in each group.
Conclusions:
We can isolate CXCR4(+) DPCs from DPCs with MACS and identify them by immunohistochemistry and flow cytometry.
