北京口腔医学
北京口腔醫學
북경구강의학
BEIJING JOURNAL OF STOMATOLOGY
2014年
5期
241-245
,共5页
DLX2%干细胞%脐带%成骨分化
DLX2%榦細胞%臍帶%成骨分化
DLX2%간세포%제대%성골분화
Distal-less homeobox 2 (DLX2)%Mesenchymal stem cells%Wharton’s jelly of the umbilical cord%Osteogenic differentiation
目的：研究Distal-less homeobox 2(DLX2)对脐带干细胞成骨定向分化能力的影响。方法成骨分化诱导培养基诱导脐带干细胞体外成骨分化；逆转录病毒转染构建过表达DLX2的脐带干细胞,进行 DLX2获得性功能研究；碱性磷酸酶活性实验检测成骨早期分化指标-碱性磷酸酶活性；茜素红染色及钙离子定量分析检测干细胞体外矿化能力；实时荧光定量反转录PCR( real time reverse transcription PCR,qRT-PCR)检测DLX2及成骨分化相关基因-Sp7转录因子、骨涎蛋白和骨钙素的表达。结果 qRT-PCR结果显示,DLX2在牙周膜干细胞的表达明显高于非牙源性干细胞-脐带干细胞、骨髓干细胞、脂肪干细胞。碱性磷酸酶活性结果显示过表达DLX2明显增强脐带干细胞的碱性磷酸酶活性；茜素红染色及钙离子定量分析结果显示过表达DLX2明显增强脐带干细胞体外矿化能力；qRT-PCR结果显示过表达DLX2明显促进Sp7转录因子、骨涎蛋白和骨钙素的表达。结论 DLX2具有促进脐带干细胞体外成骨分化的潜能。
目的：研究Distal-less homeobox 2(DLX2)對臍帶榦細胞成骨定嚮分化能力的影響。方法成骨分化誘導培養基誘導臍帶榦細胞體外成骨分化；逆轉錄病毒轉染構建過錶達DLX2的臍帶榦細胞,進行 DLX2穫得性功能研究；堿性燐痠酶活性實驗檢測成骨早期分化指標-堿性燐痠酶活性；茜素紅染色及鈣離子定量分析檢測榦細胞體外礦化能力；實時熒光定量反轉錄PCR( real time reverse transcription PCR,qRT-PCR)檢測DLX2及成骨分化相關基因-Sp7轉錄因子、骨涎蛋白和骨鈣素的錶達。結果 qRT-PCR結果顯示,DLX2在牙週膜榦細胞的錶達明顯高于非牙源性榦細胞-臍帶榦細胞、骨髓榦細胞、脂肪榦細胞。堿性燐痠酶活性結果顯示過錶達DLX2明顯增彊臍帶榦細胞的堿性燐痠酶活性；茜素紅染色及鈣離子定量分析結果顯示過錶達DLX2明顯增彊臍帶榦細胞體外礦化能力；qRT-PCR結果顯示過錶達DLX2明顯促進Sp7轉錄因子、骨涎蛋白和骨鈣素的錶達。結論 DLX2具有促進臍帶榦細胞體外成骨分化的潛能。
목적：연구Distal-less homeobox 2(DLX2)대제대간세포성골정향분화능력적영향。방법성골분화유도배양기유도제대간세포체외성골분화；역전록병독전염구건과표체DLX2적제대간세포,진행 DLX2획득성공능연구；감성린산매활성실험검측성골조기분화지표-감성린산매활성；천소홍염색급개리자정량분석검측간세포체외광화능력；실시형광정량반전록PCR( real time reverse transcription PCR,qRT-PCR)검측DLX2급성골분화상관기인-Sp7전록인자、골연단백화골개소적표체。결과 qRT-PCR결과현시,DLX2재아주막간세포적표체명현고우비아원성간세포-제대간세포、골수간세포、지방간세포。감성린산매활성결과현시과표체DLX2명현증강제대간세포적감성린산매활성；천소홍염색급개리자정량분석결과현시과표체DLX2명현증강제대간세포체외광화능력；qRT-PCR결과현시과표체DLX2명현촉진Sp7전록인자、골연단백화골개소적표체。결론 DLX2구유촉진제대간세포체외성골분화적잠능。
Objective To investigate the role of Distal-less homeobox 2 ( DLX2 ) in the osteogenic differentiation potential of mesenchymal stem cells derived from Wharton’s jelly of the umbilical cord ( WJCMSCs) . Methods Osteogenic medium was used to induce the osteogenic differentiation of WJCMSCs. Retroviral Flag-DLX2 was used to establish the stable cell and over-expressed DLX2 for Gain-of-function study in WJCMSCs. The early marker of osteogenic differentiation-ALP activity was detected by alkaline phosphatase ( ALP) activity assay. Alizarin-red staining and quantitative analysis of calcium were used to investigate the mineralization potentials of WJCMSCs in vitro. The osteogenesis related genes expressions, Sp7 transcript factor ( osterix,OSX) , bone sialoprotein ( BSP) and osteocalcin ( OCN) were examined by real time RT-PCR . Results The expression of DLX2 was higher in periodontal ligament stem cells ( PDLSCs) than in non-dental tissues’ derived stem cells( WJCMSCs, bone marrow stem cells and adipose derived stem cells) . Over-expression of DLX2 promoted ALP activity, mineralization and the expressions of OSX, BSP and OCN in WJCMSCs. Conclusion DLX2 enhanced osteogenic differentiation potential of WJCMSCs in vitro.
