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目的探讨鼠骨骼肌肌源干细胞(MDC)自体移植在压力性尿失禁治疗中的可行性。方法采用差速贴壁法分离、纯化培养6只雌性大鼠的MDC,体外脂质体介导法转染pEGFP-N1质粒入MDC,转染成功后将筛选的MDC注射于自体膀胱颈和尿道周围,5、15 d处死大鼠切取膀胱及后尿道,组织学检查观察注射组织形态学改善,免疫组化检测GFP蛋白的表达,观察MDC存活情况。结果5、15 d注射部位组织未见明显炎症改变及炎性细胞浸润,局部细胞层数增多,免疫组化显示各时间点大量细胞细胞质内可见棕黄色GFP蛋白表达,且在5、15 d两个时间点GFP阳性细胞数量和染色强度没有明显改变。结论MDC自体膀胱尿道周围注射可持续存活,自体MDC有可能成为治疗压力性尿失禁的一种理想材料。
Objective To investigate the feasibility of autologous transplantation of murine skeletal muscle myogenic stem cells (MDC) in the treatment of stress urinary incontinence. Methods MDC of 6 female rats were isolated and purified by differential adherence method. The plasmid pEGFP-N1 was transfected into MDC by liposome in vitro. MDC was screened and injected into bladder neck and urethra Surrounding rats were sacrificed on day 5 and day 15 for excision of the bladder and urethra. Histological examination of the injected tissue was observed by histological examination. The expression of GFP protein was detected by immunohistochemistry. The survival of MDC was observed. Results No inflammatory changes and inflammatory cell infiltration were found in the injection site at 5 and 15 days. The number of local cell layers increased. Immunohistochemistry showed that the expression of brown-yellow GFP protein was observed in cytoplasm at various time points. There was no significant change in the number of GFP positive cells and staining intensity at any time point. Conclusion MDC can survive continuously around the bladder and urethra. Autologous MDC may become an ideal material for the treatment of stress urinary incontinence.