华西口腔医学杂志
華西口腔醫學雜誌
화서구강의학잡지
WEST CHINA JOURNAL OF STOMATOLOGY
2015年
3期
238-243
,共6页
基质细胞衍生因子-1%牙龈干细胞%趋化作用
基質細胞衍生因子-1%牙齦榦細胞%趨化作用
기질세포연생인자-1%아간간세포%추화작용
stromal cell-derived factor-1%gingival mesenchymal stem cells%chemotaxis
目的:研究基质细胞衍生因子-1(SDF-1)受体CXCR4在人牙龈干细胞(GMSCs)上的表达及SDF-1对人GMSCs的趋化效应。方法通过有限稀释法分离并培养人GMSCs,检测其表面干细胞标志物的表达情况,测试其克隆形成率及多向分化能力,利用免疫荧光染色法检测人GMSCs上SDF-1受体CXCR4的表达,用Transwell细胞培养室检测不同质量浓度SDF-1对人GMSCs的趋化反应,光镜下计数迁移至滤膜下侧面的不同视野的细胞数。结果人GMSCs具有较高的自我更新能力,在体外呈克隆状生长,表达间充质干细胞表面标志物CD44、CD73、CD90、CD105和CD166,而造血干细胞表面标志物CD14、CD34和CD45的表达为阴性。体外诱导培养的人GMSCs能够向成骨细胞及成脂细胞分化,其克隆形成率为21.4%±2.8%。免疫荧光染色显示,人GMSCs表达SDF-1受体CXCR4。SDF-1的质量浓度为100、200?ng·mL-1时,Transwell细胞培养室中迁移的细胞数目(每高倍视野分别为189.3±4.4和164.6±4.9)显著多于空白对照组(每高倍视野47.8±2.5)(P<0.01);使用CXCR4中和抗体处理后,人GMSCs的迁移效应明显受到抑制(每高倍视野降低为29.0±2.4,P<0.01)。结论人GMSCs表达趋化因子SDF-1受体CXCR4, SDF-1对人GMSCs有趋化效应,这种趋化效应可能是通过其特异性受体CXCR4介导的。
目的:研究基質細胞衍生因子-1(SDF-1)受體CXCR4在人牙齦榦細胞(GMSCs)上的錶達及SDF-1對人GMSCs的趨化效應。方法通過有限稀釋法分離併培養人GMSCs,檢測其錶麵榦細胞標誌物的錶達情況,測試其剋隆形成率及多嚮分化能力,利用免疫熒光染色法檢測人GMSCs上SDF-1受體CXCR4的錶達,用Transwell細胞培養室檢測不同質量濃度SDF-1對人GMSCs的趨化反應,光鏡下計數遷移至濾膜下側麵的不同視野的細胞數。結果人GMSCs具有較高的自我更新能力,在體外呈剋隆狀生長,錶達間充質榦細胞錶麵標誌物CD44、CD73、CD90、CD105和CD166,而造血榦細胞錶麵標誌物CD14、CD34和CD45的錶達為陰性。體外誘導培養的人GMSCs能夠嚮成骨細胞及成脂細胞分化,其剋隆形成率為21.4%±2.8%。免疫熒光染色顯示,人GMSCs錶達SDF-1受體CXCR4。SDF-1的質量濃度為100、200?ng·mL-1時,Transwell細胞培養室中遷移的細胞數目(每高倍視野分彆為189.3±4.4和164.6±4.9)顯著多于空白對照組(每高倍視野47.8±2.5)(P<0.01);使用CXCR4中和抗體處理後,人GMSCs的遷移效應明顯受到抑製(每高倍視野降低為29.0±2.4,P<0.01)。結論人GMSCs錶達趨化因子SDF-1受體CXCR4, SDF-1對人GMSCs有趨化效應,這種趨化效應可能是通過其特異性受體CXCR4介導的。
목적:연구기질세포연생인자-1(SDF-1)수체CXCR4재인아간간세포(GMSCs)상적표체급SDF-1대인GMSCs적추화효응。방법통과유한희석법분리병배양인GMSCs,검측기표면간세포표지물적표체정황,측시기극륭형성솔급다향분화능력,이용면역형광염색법검측인GMSCs상SDF-1수체CXCR4적표체,용Transwell세포배양실검측불동질량농도SDF-1대인GMSCs적추화반응,광경하계수천이지려막하측면적불동시야적세포수。결과인GMSCs구유교고적자아경신능력,재체외정극륭상생장,표체간충질간세포표면표지물CD44、CD73、CD90、CD105화CD166,이조혈간세포표면표지물CD14、CD34화CD45적표체위음성。체외유도배양적인GMSCs능구향성골세포급성지세포분화,기극륭형성솔위21.4%±2.8%。면역형광염색현시,인GMSCs표체SDF-1수체CXCR4。SDF-1적질량농도위100、200?ng·mL-1시,Transwell세포배양실중천이적세포수목(매고배시야분별위189.3±4.4화164.6±4.9)현저다우공백대조조(매고배시야47.8±2.5)(P<0.01);사용CXCR4중화항체처리후,인GMSCs적천이효응명현수도억제(매고배시야강저위29.0±2.4,P<0.01)。결론인GMSCs표체추화인자SDF-1수체CXCR4, SDF-1대인GMSCs유추화효응,저충추화효응가능시통과기특이성수체CXCR4개도적。
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.