Wisel Sheik, Chacko Simi M, Kuppusamy M Lakshmi, Pandian Ramasamy P, Khan Mahmood, Kutala Vijay Kumar, Burry Richard W, Sun Benjamin, Kwiatkowski Pawel, Kuppusamy Periannan
Center for Biomedical Electron Paramagnetic Resonance Spectroscopy and Imaging, Division of Cardiothoracic Surgery, Department of Surgery, Davis Heart and Lung Research Institute, The Ohio State University, Columbus, OH 43210, USA.
Am J Physiol Heart Circ Physiol. 2007 Mar;292(3):H1254-61. doi: 10.1152/ajpheart.01058.2006. Epub 2006 Dec 1.
We report the labeling (internalization) of skeletal myoblasts (SMs) with a novel class of oxygen-sensing paramagnetic spin probe for noninvasive tracking and in situ monitoring of oxygenation in stem cell therapy using electron paramagnetic resonance (EPR) spectroscopy. SM cells were isolated from thigh muscle biopsies of mice and propagated in culture. Labeling of SM cells with the probe was achieved by coincubating the cells with submicron-sized (270 +/- 120 nm) particulates of the probe in culture for 48 h. The labeling had no significant effect on the viability or proliferation of the cells. The SM cells labeled with the probe were transplanted in the infarcted region of mouse hearts. The engraftment of the transplanted cells in the infarct region was verified by using MY-32 staining for skeletal myocytes. The in situ Po(2) in the heart was determined noninvasively and repeatedly for 4 wk after transplantation. The results showed significant enhancement of myocardial oxygenation at the site of cell transplant compared with untreated control. In conclusion, labeling of SM cells with the oxygen-sensing spin probe offers a unique opportunity for the noninvasive monitoring of transplanted cells as well as in situ tissue Po(2) in infarcted mouse hearts.
我们报告了使用一种新型的氧敏感顺磁自旋探针标记(内化)骨骼肌成肌细胞(SMs),以便利用电子顺磁共振(EPR)光谱对干细胞治疗中的氧合作用进行无创跟踪和原位监测。从小鼠大腿肌肉活检中分离出SM细胞并在培养中进行增殖。通过在培养中将细胞与亚微米大小(270±120 nm)的探针颗粒共孵育48小时,实现了用该探针标记SM细胞。这种标记对细胞的活力或增殖没有显著影响。用探针标记的SM细胞被移植到小鼠心脏的梗死区域。通过使用针对骨骼肌细胞的MY-32染色,验证了移植细胞在梗死区域的植入情况。在移植后4周内,对心脏中的原位氧分压(Po(2))进行了无创且重复的测定。结果显示,与未处理的对照组相比,细胞移植部位的心肌氧合有显著增强。总之,用氧敏感自旋探针标记SM细胞为无创监测移植细胞以及梗死小鼠心脏中的原位组织Po(2)提供了独特的机会。