摘要
目的探讨 CM-DiI 标记小鼠骨骼肌卫星细胞后移植治疗心肌梗死的体外体内示踪作用。方法 CM-DiI 标记提纯后小鼠骨骼肌卫星细胞,检测标记率及荧光情况,观察 CM-DiI 对细胞增殖分化的影响。小鼠心梗模型心肌内注射标记后的卫星细胞,于移植后 1、3、7、14、21、28 天取出心脏行冰冻切片,观察移植细胞在体存活分化情况。结果 CM-DiI 标记骨骼肌卫星细胞效率达 (93.3±0.95)%,3 代后仍保持红色荧光,对细胞增殖及分化无明显影响。标记细胞移植 1 天后即可见细胞团聚,3 天后细胞排列整齐,4 周后仍可见移植细胞。结论 CM-DiI 细胞标记液标记骨骼肌卫星细胞,安全简单,并可示踪移植后细胞的存活分化情况。
Object ive To investigate the tracing effect of CM-DiI as a cell t racer for skeletal myoblasts transplantation in mous emyocardiac infarct ion models. Method: Skeletal myoblasts，which were isolated and puri��ed from C57/BL6 mouse, were labled with CM-DiI. Label ing ef��ciency was evaluated by ��owcytometer and ��uorescence micros cope. The proli feration and di fferentiation of CM-DiI labeled myoblsts were observed by cell IQ system. After the labeled skeletal myoblasts were transplanted into is chemic myocardium by injection, heart samples were then taken for frozen sect ions at various t ime points (day-1, day-3, day-7, day-14, day-21 and day-28) after t ransplantation. The survival and differentiation of grafted cells in vivo were observed under ��uorescence microscope. Results: The labeling ef��ciency of CM-Di I was (93.3±0.95)%. Fluores cence of labled cells could be maintained after 3 generat ions. There were no in��uence on the proliferation and differentiation of skeletal myobls ts after labeling. In vivo, the labled cells aggregated 24 hours after transplantation. Grafted cells were seen to be aligned and well oriented at day-3. And ��uorescence of grafted cells could s till be observed after 28 days. Conclus ion: The method of label ing skeletal myoblas ts with CM-DiI was effective and easy, and could t race the survival and different iat ion of grafted cells in vivo.
出处
《中国分子心脏病学杂志》
CAS
2011年第5期285-289,共5页
Molecular Cardiology of China
基金
上海市浦江人才计划(10PJ1402000)
教育部新教师基金(20090071120032)
