Culturing and characterization of human gingival mesenchymal stem cells and their chemotactic responses to stromal cell-derived factor-1
10.7518/hxkq.2015.03.005
- VernacularTitle:人牙龈干细胞的分离鉴定及基质细胞衍生因子-1对其趋化效应的研究
- Author:
Lingqian DU
1
;
Pishan YANG
;
Shaohua GE
Author Information
1. 山东大学第二医院口腔科
- Keywords:
stromal cell-derived factor-1;
gingival mesenchymal stem cells;
chemotaxis
- From:
West China Journal of Stomatology
2015;(3):238-243
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo?investigate?the?expression?of?chemokine?stromal?cell-derived?factor-1?(SDF-1)?receptor?CXCR4?in?human?gingival?mesenchymal?stem?cells?(GMSCs)?and?the?migration?potential?of?GMSCs?stimulated?with?SDF-1.?Methods?Human?GMSCs?were?isolated?by?single-cell?cloning?method.?Their?cell?surface?markers?were?characterized?by?flow?cytometry,?and?the?rate?of?colony?formation?was?evaluated.?Differentiation?assay?was?used?to?detect?the?differentiation?potential?of?GMSCs.?The?expression?of?chemokine?SDF-1?receptor?CXCR4?in?GMSCs?was?detected?by?immunocytochemical?staining.?The?che-motactic?effect?of?SDF-1?on?GMSCs?was?detected?using?a?24-multiwell?Transwell?cell?culture?chamber.?The?number?of?net?migrated?cells?was?counted?in?different?microscope?fields.?Results???Human?GMSCs?possessed?high?self-renewal?potential?and?formed?single-cell?colonies?cultured?in vitro.?GMSCs?expressed?mesenchymal?stem?cells-associated?markers?CD44,?CD73,?CD90,?CD105,?and?CD166,?and?the?expression?of?hemopoietic?stem?cell?surface?markers?CD14,?CD34,?and?CD45?was?negative.?GMSCs?differentiated?into?osteoblasts?and?adipocytes?under?defined?culture?conditions.?The?colony?forming?unit-fibroblastic?for?GMSCs?was?21.4%±2.8%.?Immunocytochemical?staining?demonstrated?that?GMSCs?expressed?chemokine?SDF-1?receptor?CXCR4.?The?number?of?GMSCs?migrating?at?concentrations?of?100?ng·mL-1?and?200?ng·mL-1?of?SDF-1?in?the?Transwell?cell?culture?chamber?was?significantly?higher?than?that?of?the?negative?control?(189.3±4.4,?164.6±4.9?cells/field?vs.?47.8±2.5?cells/field,?P<0.01).?Treatment?with?the?CXCR4?neutralizing?antibody,?an?antagonist?for?CXCR4,?significantly?reduced?the?migratory?effect?compared?with?the?negative?con-trols?(29.0±2.4?cells/field?vs.?47.8±2.5?cells/field,?P<0.01).?Conclusion???Human?GMSCs?express?chemokine?SDF-1?receptor?CXCR4.?SDF-1?may?participate?in?regulating?chemotaxis?of?human?GMSCs.?Results?suggest?that?the?migration?induced?by?SDF-1?is?mediated?by?CXCR4.
