中国组织工程研究
中國組織工程研究
중국조직공정연구
Journal of Clinical Rehabilitative Tissue Engineering Research
2013年
45期
7885-7890
,共6页
干细胞%脐带脐血干细胞%脐带%间充质干细胞%碱性成纤维细胞生长因子%基因转染%腺病毒%干细胞图片文章
榦細胞%臍帶臍血榦細胞%臍帶%間充質榦細胞%堿性成纖維細胞生長因子%基因轉染%腺病毒%榦細胞圖片文章
간세포%제대제혈간세포%제대%간충질간세포%감성성섬유세포생장인자%기인전염%선병독%간세포도편문장
背景：目前多采取重组腺相关病毒作为载体介导碱性成纤维细胞生长因子基因转染的方法，将外源性碱性成纤维细胞生长因子基因转入脐带间充质干细胞内并持续表达，调控细胞增殖及定向分化，取得高效持久的治疗作用。<br> 目的：了解采用重组腺相关病毒作为载体介导碱性成纤维细胞生长因子基因转染对体外培养的脐带间充质干细胞增殖及细胞周期的影响。<br> 方法：体外培养脐带间充质干细胞，经重组腺相关病毒作为载体介导碱性成纤维细胞生长因子基因转染，分为对照组、空载病毒组、碱性成纤维细胞生长因子转染组。用RT-PCR，Western blot检测脐带间充质干细胞在转染前后碱性成纤维细胞生长因子基因和蛋白的表达。应用细胞生长曲线、CCK-8观察细胞生长的优化作用，采用流式细胞术测定细胞周期分布的变化。<br> 结果与结论：转染碱性成纤维细胞生长因子后，转染组与对照组、空载病毒组相比碱性成纤维细胞生长因子基因和蛋白水平均有表达，细胞的生长速度明显增快，细胞周期 G0/G1期减少，S 期细胞数增多，各组间差异有显著性意义(P<0.05)。说明，通过重组腺相关病毒作为载体介导碱性成纤维细胞生长因子基因转染能促进体外培养的脐带间充质干细胞增殖，对其培养有优化作用。
揹景：目前多採取重組腺相關病毒作為載體介導堿性成纖維細胞生長因子基因轉染的方法，將外源性堿性成纖維細胞生長因子基因轉入臍帶間充質榦細胞內併持續錶達，調控細胞增殖及定嚮分化，取得高效持久的治療作用。<br> 目的：瞭解採用重組腺相關病毒作為載體介導堿性成纖維細胞生長因子基因轉染對體外培養的臍帶間充質榦細胞增殖及細胞週期的影響。<br> 方法：體外培養臍帶間充質榦細胞，經重組腺相關病毒作為載體介導堿性成纖維細胞生長因子基因轉染，分為對照組、空載病毒組、堿性成纖維細胞生長因子轉染組。用RT-PCR，Western blot檢測臍帶間充質榦細胞在轉染前後堿性成纖維細胞生長因子基因和蛋白的錶達。應用細胞生長麯線、CCK-8觀察細胞生長的優化作用，採用流式細胞術測定細胞週期分佈的變化。<br> 結果與結論：轉染堿性成纖維細胞生長因子後，轉染組與對照組、空載病毒組相比堿性成纖維細胞生長因子基因和蛋白水平均有錶達，細胞的生長速度明顯增快，細胞週期 G0/G1期減少，S 期細胞數增多，各組間差異有顯著性意義(P<0.05)。說明，通過重組腺相關病毒作為載體介導堿性成纖維細胞生長因子基因轉染能促進體外培養的臍帶間充質榦細胞增殖，對其培養有優化作用。
배경：목전다채취중조선상관병독작위재체개도감성성섬유세포생장인자기인전염적방법，장외원성감성성섬유세포생장인자기인전입제대간충질간세포내병지속표체，조공세포증식급정향분화，취득고효지구적치료작용。<br> 목적：료해채용중조선상관병독작위재체개도감성성섬유세포생장인자기인전염대체외배양적제대간충질간세포증식급세포주기적영향。<br> 방법：체외배양제대간충질간세포，경중조선상관병독작위재체개도감성성섬유세포생장인자기인전염，분위대조조、공재병독조、감성성섬유세포생장인자전염조。용RT-PCR，Western blot검측제대간충질간세포재전염전후감성성섬유세포생장인자기인화단백적표체。응용세포생장곡선、CCK-8관찰세포생장적우화작용，채용류식세포술측정세포주기분포적변화。<br> 결과여결론：전염감성성섬유세포생장인자후，전염조여대조조、공재병독조상비감성성섬유세포생장인자기인화단백수평균유표체，세포적생장속도명현증쾌，세포주기 G0/G1기감소，S 기세포수증다，각조간차이유현저성의의(P<0.05)。설명，통과중조선상관병독작위재체개도감성성섬유세포생장인자기인전염능촉진체외배양적제대간충질간세포증식，대기배양유우화작용。
BACKGROUND:At present, exogenous basic fibroblast growth factor gene can be transfected into umbilical cord mesenchymal stem cells via a recombinant adeno-associated virus vector and exhibit sustained expression in transfected cells. This method can regulate cellproliferation and directed differentiation to obtain efficient long-lasting therapeutic effects. <br> OBJECTIVE:To investigate the effects of basic fibroblast growth factor gene transfection via a recombinant adeno-associated virus vector on the proliferation and cellcycle of human umbilical cord mesenchymal stem cells cultured in vitro. <br> METHODS:Human umbilical cord mesenchymal stem cells were cultured by the suspension culture in vitro, and were transfected by recombinant adeno-associated virus-mediated basic fibroblast growth factor gene. Cultured cells were divided into three groups:control group, basic fibroblast growth factor group, and recombinant adeno-associated virus group. Reverse transcription-PCR and western blot were used to assess the knockdown efficiency. cellular proliferation was determined by cellgrowth curve and cellCounting Kit-8 assay. The cellcycle was analyzed by flow cytometry. <br> RESULTS AND CONCLUSION:Compared with the other two groups, the expression of basic fibroblast growth factor mRNA and protein increased significantly, the cellgrowth speed was also significantly increased, the cellcycle of G0/G1 phase was decreased and cellnumber in S phase was increased in the basic fibroblast growth factor group after transfection. These findings suggest that the recombinant adeno-associated virus-mediated basic fibroblast growth factor gene can promote the proliferation of umbilical cord mesenchymal stem cells proliferation cultured in vitro, and also can optimize the cellculture.
