骨髓单个核细胞经冠状动脉移植治疗心肌梗死的实验研究

Experimental study on treatment of myocardial infarction by intracoronary transplantation of isogenic bone marrow mononuclear cells

  • 摘要: 目的: 观察同基因骨髓单个核细胞经冠状动脉植入心脏后的分布及分化状况。方法: 用雄性近交系Lew is大鼠作为模型动物。首先,在大鼠颈部异位心脏移植模型基础上,以冷冻法制备急性心肌梗死模型。其次,心肌梗死模型建立2 d后,分离、纯化大鼠骨髓单个核细胞,并以PKH26红色荧光染料标记。然后,经移植心脏冠状动脉输注PKH26标记的同基因骨髓单个核细胞(实验组)和DMEM培养液(对照组)。最后,细胞移植7 d后,采集各标本,检测植入细胞分布及分化状况。结果: 病理检查证实冷冻区域心肌变性坏死且界线清楚。利用该模型成功的模拟了临床经冠状动脉输注自体骨髓单个核细胞的治疗方法。PKH26红色荧光阳性细胞主要集中于实验组大鼠移植心脏的冷冻梗死区,脾脏、骨髓也可见少量阳性细胞分布,移植心脏的非梗死区及其余脏器组织切片均未见红色荧光阳性细胞;梗死区内PKH26标记的细胞部分表达CD34;梗死区内connexin43的表达明显高于对照组。结论: 供者同基因骨髓单个核细胞经冠状动脉移植后,能够存活并定向迁移至心肌损伤区域,并且可能促使梗死区出现心肌再生。

     

    Abstract: Objective: To investigate the feasibility of intracoronary administration of isogenic bone marrow mononuclear cells(BMMNCs) by assessing the fate of transplanted,purified,labeled cells in ectopic transplanted cryo-damaged hearts in rats. Methods: Inbred isogenic adult(8-10 weeks old) Lewis rats were used as donors and recipients.First,myocardial damage was obtained in the hearts of donor rats by placing a frozen metal rod on the anterior left ventricular wall for 20 seconds.Second,the injured hearts were procured and then isogenic heterotopic heart transplantation were performed in the cervical area of recipient rats.Third,Two days after the injury,isogenic BMMNCs were purified and labeled with PKH26(a red fluorescent cell dye) and approximate 2×107 cells were infused through the coronary artery of infarcted transplanted hearts of the experimental group,while the control group were injected with DMEM.Finally,seven days after the infusion,the infarcted and normal hearts,lungs,livers,kidneys,spleens and bone marrow were harvested from recipient rats to track the fate of transplanted,labeled BMMNCs. Results: The myocardial degeneration and necrosis with clear border were identified in the cryo-injured heart of model rats by pathology.This model was successfully used to mimic intracoronary transplantation of autologous bone marrow mononuclear cells in clinical setting.Labeled cells were found only in the injured myocardium of treated animals and not in the normal tissue,and a limited number of cells were identified in the spleen and bone marrow of the treated recipients.Some of the labeled cells in the infarcted area were CD34 staining positive.Expression of connexin43 in the infarcted area of treated rats was significantly higher than that in the infarcted area of controls. Conclusion: Our data suggest that intracoronarily injected BMMNCs can survive and traffic through the circulation to the site of damaged myocardium and may lead to regeneration of new myocardium.

     

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