Sheikh Ahmad Y, Lin Shu-An, Cao Feng, Cao Yuan, van der Bogt Koen E A, Chu Pauline, Chang Ching-Pin, Contag Christopher H, Robbins Robert C, Wu Joseph C
Department of Cardiothoracic Surgery, Stanford University School of Medicine, Edwards Building R354, Stanford, California 94305-5344, USA.
Stem Cells. 2007 Oct;25(10):2677-84. doi: 10.1634/stemcells.2007-0041. Epub 2007 Jul 12.
Bone marrow mononuclear cell (BMMC) therapy shows promise as a treatment for ischemic heart disease. However, the ability to monitor long-term cell fate remains limited. We hypothesized that molecular imaging could be used to track stem cell homing and survival after myocardial ischemia-reperfusion (I/R) injury. We first harvested donor BMMCs from adult male L2G85 transgenic mice constitutively expressing both firefly luciferase (Fluc) and enhanced green fluorescence protein reporter gene. Fluorescence-activated cell sorting analysis revealed approximately 0.07% of the population to consist of classic hematopoietic stem cells (lin-, thy-int, c-kit+, Sca-1+). Afterward, adult female FVB recipients (n = 38) were randomized to sham surgery or acute I/R injury. Animals in the sham (n = 16) and I/R (n = 22) groups received 5 x 10(6) of the L2G85-derived BMMCs via tail vein injection. Bioluminescence imaging (BLI) was used to track cell migration and survival in vivo for 4 weeks. BLI showed preferential homing of BMMCs to hearts with I/R injury compared with sham hearts within the first week following cell injection. Ex vivo analysis of explanted hearts by histology confirmed BLI imaging results, and quantitative real-time polymerase chain reaction (for the male Sry gene) further demonstrated a greater number of BMMCs in hearts with I/R injury compared with the sham group. Functional evaluation by echocardiography demonstrated a trend toward improved left ventricular fractional shortening in animals receiving BMMCs. Taken together, these data demonstrate that molecular imaging can be used to successfully track BMMC therapy in murine models of heart disease. Specifically, we have demonstrated that systemically delivered BMMCs preferentially home to and are retained by injured myocardium. Disclosure of potential conflicts of interest is found at the end of this article.
骨髓单个核细胞(BMMC)疗法有望成为治疗缺血性心脏病的方法。然而,监测细胞长期命运的能力仍然有限。我们推测分子成像可用于追踪心肌缺血再灌注(I/R)损伤后干细胞的归巢和存活情况。我们首先从成年雄性L2G85转基因小鼠中获取供体BMMC,这些小鼠组成性表达萤火虫荧光素酶(Fluc)和增强型绿色荧光蛋白报告基因。荧光激活细胞分选分析显示,约0.07%的细胞群体为经典造血干细胞(lin-、thy-int、c-kit+、Sca-1+)。随后,成年雌性FVB受体(n = 38)被随机分为假手术组或急性I/R损伤组。假手术组(n = 16)和I/R组(n = 22)的动物通过尾静脉注射接受5×10⁶个源自L2G85的BMMC。生物发光成像(BLI)用于在体内追踪细胞迁移和存活4周。BLI显示,与假手术组心脏相比,在细胞注射后的第一周内,BMMC更倾向于归巢到I/R损伤的心脏。通过组织学对取出的心脏进行体外分析证实了BLI成像结果,定量实时聚合酶链反应(针对雄性Sry基因)进一步表明,与假手术组相比,I/R损伤心脏中的BMMC数量更多。通过超声心动图进行的功能评估表明,接受BMMC的动物左心室缩短分数有改善的趋势。综上所述,这些数据表明分子成像可成功追踪心脏病小鼠模型中的BMMC治疗。具体而言,我们已经证明全身递送的BMMC优先归巢到受损心肌并在其中保留。潜在利益冲突的披露见本文末尾。