实时定量PCR法检测骨髓间充质干细胞移植到梗死心肌后的存活数量_临床医学论文.docVIP

实时定量PCR法检测骨髓间充质干细胞移植到梗死心肌后的存活数量_临床医学论文 实时定量PCR法检测骨髓间充质干细胞移植到梗死心肌后的存活数量_临床医学论文 作者：王业焕，张浩，袁昕，侯剑锋，刘学彬，叶珏，孙成超 【摘要】 目的 ：建立一种干细胞移植到梗死心肌后检测其存活数量的有效方法。方法 ：选取10只雌性SD大鼠结扎前降支，心肌梗死3周后，随机分成移植组与对照组，同时选取雄性大鼠培养出骨髓间充质干细胞(MSCs)，分别将MSCs(3×106个，50μL)及等体积的PBS液移植到梗死心肌中，24 h后通过实时定量PCR(Real-time PCR)检测雄性大鼠Y染色体上特有的基因SRY，从而得知移植MSCs的存活数量。结果：MSCs移植24 h后，移植组梗死心肌中检测到SRY基因数占移植总数的14.9%±8.3%，存活细胞的绝对数量为447 195±248 090，而对照组均未检测到SRY基因的存在。结论：用Real-time PCR的方法能有效地检测到具有SRY基因的干细胞，从而能准确地得知移植干细胞的存活数量，Real-time PCR技术在心梗后干细胞移植的基础研究及临床应用中具有重要的价值。 【关键词】 实时定量PCR;心肌梗死;细胞移植 　　Abstract: Objective: To establish an effective method to detect surviving number of the stem cells in vivo. Methods: Ten adult female SD rats were randomly assigned into two groups: the bone marrow derived from mesenchymal stem cells (MSCs) transplantion group and the control group. Anterior myocardial infarction was induced by ligation of the proximal left anterior descending artery. Using an insulin syringe， PBS (50μL) and MSC (3×106/50μL) of male rat sources were injected into infarcted area in transplantation group and control group，respectively. To evaluate the viability of MSCs injected into infarcted area， Real-time PCR was used to detect the SRY gene which is a specific marker of Y chromosome in 24 hours after MSC transplantation. Results: Twenty four hours after MSC transplantation， the cells were detected in the infarcted myocardium in six rats of transplantation group. The number of surviving cells was 447 195±248 090 ;the number of detected SRY to the number of transplanted cells was 14.9%±8.3%. In contrast，no sry gene were detected in control group. Conclusion: Real-time PCR is a sensitive and accurate method to detect the viability of transplanted stem cells in infarcted myocardium， and this method is valuable for both the laboratory study and clinical practice in the field of stem cell therapy for myocardial infarction. 　　Key words: Real-time PCR;myocardial infarction;cell transplantation 　　心肌梗死后，心肌细胞可出现永久性的丢失，且成熟心肌细胞不可再生，现有治疗