Tracing of GFP-labeled stem cells transplanted to discs for treating degenerative discs
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摘要: 目的 探讨绿色荧光蛋白(GFP)示踪时效及GFP标记的骨髓间充质干细胞(MSCs)在移植后参与退变修复过程中的细胞增殖情况.方法 使用针吸髓核法建立兔的椎间盘退变模型,体外培养MSCs,并用含有绿色荧光蛋白标记的腺病毒载体(Adeno-XTM-GFP)转染,成为GFP标记的MSCs(GFP-MSCs),于造模2周后植入造模的椎间盘内.在移植后第2、4、6、8周用激光共聚焦显微成像技术,观察MSCs的增殖情况,对比GFP本身荧光和免疫荧光的示踪时间.结果 非免疫荧光组GFP本身荧光强度能够持续到4周,后较微弱甚至难以辨别;免疫荧光组用FITC荧光二抗标记的GFP-MSCs在8周内都能够持续稳定的显现荧光,4个时间点GFP-MSCs的阳性率的可信区间分别是:2周(14.64±2.05)%;4周(21.85±2.45)%;6周(31.03±4.03)%;8周(36.37±4.42)%.结论 GFP-MSCs在椎间盘微环境中能够稳定增殖.若示踪研究需要持续超过4周,则虚对GFP行免疫荧光染色,可以使有效示踪时间持续至少8周.
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