Abstract
The purpose of this study was to investigate and compare the feasibility of rat sodium iodide symporter (rNIS) and human sodium iodide symporter (hNIS) as reporter genes for noninvasive monitoring of rat bone marrow mesenchymal stem cells (rBMSCs) transplanted into infarcted rat myocardium. rBMSCs were isolated from rat bone marrow. Adenovirus (Ad) was reconstructed to contain rNIS-enhanced green fluorescent protein (eGFP) or hNIS-eGFP. The transfection efficiency of Ad/eGFP/rNIS and Ad/eGFP/hNIS to rBMSCs was measured by real-time polymerase chain reaction, flow cytometry, Western blot, and immunofluorescence staining. The transfected rBMSCs were transplanted into infarcted rat myocardium followed by a single-photon emission computed tomography (SPECT) study with (99m)Tc-pertechnetate as the radiotracer and by autoradiography. The isolated rBMSCs were CD29, CD44, and CD90 positive and CD34, CD45, and CD11b negative. The expression of rNIS and hNIS in the transfected rBMSCs at both gene and protein levels was obviously higher than that without transfection. The myocardium of rats transplanted with transfected rBMSCs could be visualized by SPECT owing to the accumulation of (99m)Tc-pertechnetate in rBMSCs mediated by exogenous NIS genes. The accumulation of (99m)Tc-pertechnetate in myocardium mediated by rNIS was higher than that by hNIS, which was also confirmed by autoradiography. Both rNIS and hNIS are useful reporter genes to monitor BMSCs transplanted into infarcted myocardium in vivo with rNIS being superior to hNIS as the reporter gene.
Highlights
W ITH INCREASED STANDARD OF LIVING, myocardium infarction becomes one of the diseases with the highest attack rates and mortality
We investigated the feasibility of rat sodium iodide symporter and human sodium iodide symporter as reporter genes to monitor the fate of rat bone mesenchymal stem cells transplanted into infarcted rat myocardium by using single-photon emission computed tomography (SPECT) and 99mTcO42
Based on the average of three fluorescence-activated cell sorter (FACS) analyses, rat bone marrow mesenchymal stem cells (rBMSCs) were uniformly positive for extracellular matrix protein CD90 (98.07 6 5.62%), b1 integrin CD29 (95.96 6 2.80%), and hyaluronate receptor CD44 (96.15 6 3.74%); a low to negligible level of hematopoietic progenitor cell marker CD34 (0.09 6 0.02%); integrin aM chain CD11b (0.21 6 0.05%); and leukocyte common antigen CD45 (0.49 6 0.15%)
Summary
W ITH INCREASED STANDARD OF LIVING, myocardium infarction becomes one of the diseases with the highest attack rates and mortality. Stem cell transplantation is an effective means of treating infarcted myocardium. Bone mesenchymal stem cells (BMSCs) have been found to stimulate myocardial repair or actual regeneration because of the ability to expand many logfold in vitro and unique immune characteristics allowing their use as an allogeneic graft.[1,2,3] To promote the effect of repairing infarcted myocardium, BMSCs have been modified in many different ways. In the process of curing myocardial infarction, it is very important to find a noninvasive method to monitor the transplanted BMSCs at real time in vivo. There are three primary methods to track transplanted BMSCs in vivo, namely, magnetic resonance imaging (MRI),[12,13,14] optical imaging,[15,16,17,18] and radionuclide imaging.[19,20,21].
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