中国组织工程研究
中國組織工程研究
중국조직공정연구
Journal of Clinical Rehabilitative Tissue Engineering Research
2015年
14期
2250-2254
,共5页
干细胞%培养%端粒酶反转录酶%许旺细胞%基因转染%大鼠%细胞增殖%细胞周期
榦細胞%培養%耑粒酶反轉錄酶%許旺細胞%基因轉染%大鼠%細胞增殖%細胞週期
간세포%배양%단립매반전록매%허왕세포%기인전염%대서%세포증식%세포주기
Telomerase%Schwann Cells%Genes%Transfection
背景：研究表明，基因修饰许旺细胞可使许旺细胞在体内存活时间延长，促进神经再生和功能的恢复。目的：以反转录病毒 PLXSN 为载体，将 hTERT 基因转染入体外培养的大鼠许旺细胞，检测许旺细胞端粒酶活性及细胞生物学特性。方法：体外培养Wistar大鼠许旺细胞，经反转录病毒PLXSN为载体介导人端粒酶反转录酶基因转染，在同等条件下进行空载病毒转染，以正常培养的许旺细胞为对照组。采用RT-PCR，Western blot检测许旺细胞人端粒酶反转录酶基因和蛋白的表达，流式细胞仪测定细胞周期分布的变化。以细胞生长曲线、MTT比色法观察细胞生长的优化作用。结果与结论：人端粒酶反转录酶基因转染许旺细胞48 h后，检测到人端粒酶反转录酶mRNA和蛋白水平表达明显。与对照组和空载病毒组比较，细胞的生长速度明显增快，G 0/G 1期细胞数减少，S期细胞数增多，差异有显著性意义(P<0.05)。结果表明通过反转录病毒PLXSN 为载体介导人端粒酶反转录酶基因转染使许旺细胞端粒酶活性明显升高，能够促进体外培养的大鼠许旺细胞增殖。
揹景：研究錶明，基因脩飾許旺細胞可使許旺細胞在體內存活時間延長，促進神經再生和功能的恢複。目的：以反轉錄病毒 PLXSN 為載體，將 hTERT 基因轉染入體外培養的大鼠許旺細胞，檢測許旺細胞耑粒酶活性及細胞生物學特性。方法：體外培養Wistar大鼠許旺細胞，經反轉錄病毒PLXSN為載體介導人耑粒酶反轉錄酶基因轉染，在同等條件下進行空載病毒轉染，以正常培養的許旺細胞為對照組。採用RT-PCR，Western blot檢測許旺細胞人耑粒酶反轉錄酶基因和蛋白的錶達，流式細胞儀測定細胞週期分佈的變化。以細胞生長麯線、MTT比色法觀察細胞生長的優化作用。結果與結論：人耑粒酶反轉錄酶基因轉染許旺細胞48 h後，檢測到人耑粒酶反轉錄酶mRNA和蛋白水平錶達明顯。與對照組和空載病毒組比較，細胞的生長速度明顯增快，G 0/G 1期細胞數減少，S期細胞數增多，差異有顯著性意義(P<0.05)。結果錶明通過反轉錄病毒PLXSN 為載體介導人耑粒酶反轉錄酶基因轉染使許旺細胞耑粒酶活性明顯升高，能夠促進體外培養的大鼠許旺細胞增殖。
배경：연구표명，기인수식허왕세포가사허왕세포재체내존활시간연장，촉진신경재생화공능적회복。목적：이반전록병독 PLXSN 위재체，장 hTERT 기인전염입체외배양적대서허왕세포，검측허왕세포단립매활성급세포생물학특성。방법：체외배양Wistar대서허왕세포，경반전록병독PLXSN위재체개도인단립매반전록매기인전염，재동등조건하진행공재병독전염，이정상배양적허왕세포위대조조。채용RT-PCR，Western blot검측허왕세포인단립매반전록매기인화단백적표체，류식세포의측정세포주기분포적변화。이세포생장곡선、MTT비색법관찰세포생장적우화작용。결과여결론：인단립매반전록매기인전염허왕세포48 h후，검측도인단립매반전록매mRNA화단백수평표체명현。여대조조화공재병독조비교，세포적생장속도명현증쾌，G 0/G 1기세포수감소，S기세포수증다，차이유현저성의의(P<0.05)。결과표명통과반전록병독PLXSN 위재체개도인단립매반전록매기인전염사허왕세포단립매활성명현승고，능구촉진체외배양적대서허왕세포증식。
BACKGROUND:Studies have shown that genetical y modified Schwann cel s can survive for a longer time in vivo, and promote nerve regeneration and functional recovery. OBJECTIVE:To transfect human telomerase reverse transcriptase (hTERT) gene into rat Schwann cel s cultured in vitro via PLXSN vector, and to detect the telomerase activity and biological characteristics of Schwann cel s. METHODS:Schwann cel s from Wistar rats were cultured in vitro and transfected by PLXSN vector with (hTERT group) or without hTERT (empty vector group). Normal Schwann cel s were selected as control group. RT-PCR and western blot methods were used to detect the hTERT protein and mRNA levels in Schwann cel s, and flow cytometry was used to measure the cel cycle distribution. Cel growth was observed by cel growth curve and MTT colorimetric method. RESULTS AND CONCLUSION:At 48 hours after transfection, the mRNA and protein expressions of hTERT were remarkably seen in Schwann cel s. Compared with the control and empty vector groups, the cel s grew faster, the number of cel s at G 0/G 1 Schwann cel s cultured in vitro. phase was reduced, but the number of S phase cel s was increased in the hTERT group (P<0.05). These findings indicate that PLXSN vector-mediated hTERT transfection of Schwann cel s can significantly improve the activity of telomerase in Schwann cel s as wel as promote the proliferation of Schwann cells cultured in vitro.
