Long-term MRI tracking of dual-labeled adipose-derived stem cells homing into mouse carotid artery injury

Jin-Bao Qin1 Department of Vascular Surgery, Shanghai Ninth People’s Hospital Affiliated to Shanghai Jiao Tong University, School of Medicine, Shanghai;5 Vascular Center, Shanghai Jiao Tong University, Shanghai, China

Kang-An Li2 Department of Radiology, Shanghai First People’s Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai

Xiang-Xiang Li1 Department of Vascular Surgery, Shanghai Ninth People’s Hospital Affiliated to Shanghai Jiao Tong University, School of Medicine, Shanghai;5 Vascular Center, Shanghai Jiao Tong University, Shanghai, China

Qing-Song Xie3 Department of Neurosurgery, Cixi Municipal People’s Hospital, Zhejiang Province, China

Jia-Ying Lin4 Clinic for Gynecology, Charite-Universitatsmedizin Berlin, Berlin, Germany

Kai-Chuang Ye1 Department of Vascular Surgery, Shanghai Ninth People’s Hospital Affiliated to Shanghai Jiao Tong University, School of Medicine, Shanghai;5 Vascular Center, Shanghai Jiao Tong University, Shanghai, China

Mi-Er Jiang1 Department of Vascular Surgery, Shanghai Ninth People’s Hospital Affiliated to Shanghai Jiao Tong University, School of Medicine, Shanghai;5 Vascular Center, Shanghai Jiao Tong University, Shanghai, China

Gui-Xiang Zhang2 Department of Radiology, Shanghai First People’s Hospital, Shanghai Jiao Tong University School of Medicine, ShanghaiCorrespondence[email protected] [email protected]

Xin-Wu Lu1 Department of Vascular Surgery, Shanghai Ninth People’s Hospital Affiliated to Shanghai Jiao Tong University, School of Medicine, Shanghai;5 Vascular Center, Shanghai Jiao Tong University, Shanghai, ChinaCorrespondence[email protected] [email protected]

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Abstract

Background

Stem cell therapy has shown great promise for regenerative repair of injured or diseased tissues. Adipose-derived stem cells (ADSCs) have become increasingly attractive candidates for cellular therapy. Magnetic resonance imaging has been proven to be effective in tracking magnetic-labeled cells and evaluating their clinical relevance after cell transplantation. This study investigated the feasibility of imaging green fluorescent protein-expressing ADSCs (GFP-ADSCs) labeled with superparamagnetic iron oxide particles, and tracked them in vivo with noninvasive magnetic resonance imaging after cell transplantation in a model of mouse carotid artery injury.

Methods

GFP-ADSCs were isolated from the adipose tissues of GFP mice and labeled with superparamagnetic iron oxide particles. Intracellular stability, proliferation, and viability of the labeled cells were evaluated in vitro. Next, the cells were transplanted into a mouse carotid artery injury model. Clinical 3 T magnetic resonance imaging was performed immediately before and 1, 3, 7, 14, 21, and 30 days after cell transplantation. Prussian blue staining and histological analysis were performed 7 and 30 days after transplantation.

Results

GFP-ADSCs were found to be efficiently labeled with superparamagnetic iron oxide particles, with no effect on viability and proliferation. Homing of the labeled cells into the injured carotid artery tissue could be monitored by magnetic resonance imaging.

Conclusion

Magnetically labeled ADSCs with expression of GFP can home into sites of vascular injury, and may provide new insights into understanding of cell-based therapy for cardiovascular lesions.

Keywords:

Acknowledgments

This research was supported by the National Natural Science Foundation of China (30772104), the Natural Science Foundation of Shanghai Science and Technology Committee (11ZR1429300), the Medical Guiding Program of Shanghai Science and Technology Committee (1141190800), the Songjiang Medical Climbing Program, Shanghai, China (2011PD04), and the Ningbo Natural Science Foundation (2008A610093).

The authors appreciated the kind support of Professor Zun-li Shen, Yu-qing Jin, De-min Yin, Li-juan Zong, Juan-juan Wu, Jing-jun Chen, and Wan-yao Xia.

Disclosure

The authors report no conflicts of interest in this work.

Supplementary figures

Figure S1 Morphology of GFP-ADSCs labeled with 25μg/mL SPIO.

Notes: (A) Bright field of cells labeled with 25μg/mL SPIO. (B) Fluorescent microscopy image of the same view of A showed GFP expression of whole labeled cells. (C) Negative control cells without treatment with SPIO. (D) The same view of C was observed under fluorescence microscopy. Scale bar measures 100 μm.

Abbreviations: GFP-ADSCs, green fluorescent protein-adipose-derived stem cells; BF, bright field; FL, fluorescent light.

Figure S1 Morphology of GFP-ADSCs labeled with 25μg/mL SPIO.Notes: (A) Bright field of cells labeled with 25μg/mL SPIO. (B) Fluorescent microscopy image of the same view of A showed GFP expression of whole labeled cells. (C) Negative control cells without treatment with SPIO. (D) The same view of C was observed under fluorescence microscopy. Scale bar measures 100 μm.Abbreviations: GFP-ADSCs, green fluorescent protein-adipose-derived stem cells; BF, bright field; FL, fluorescent light.

Figure S2 Annexin V staining analysis of apoptosis in GFP-ADSCs labeled with 25 μg/ml SPIO.

Notes: (A) Negative control cells without treatment with SPIO. (B) GFP-ADSCs labeled with 25 μg/mL SPIO.

Abbreviation: GFP-ADSCs, green fluorescent protein adipose-derived stem cells.

Figure S2 Annexin V staining analysis of apoptosis in GFP-ADSCs labeled with 25 μg/ml SPIO.Notes: (A) Negative control cells without treatment with SPIO. (B) GFP-ADSCs labeled with 25 μg/mL SPIO.Abbreviation: GFP-ADSCs, green fluorescent protein adipose-derived stem cells.

Figure S3 Cell cycle of labeled or unlabeled GFP-ADSCs with 25 μg/ml SPIO.

Notes: FCM showed that about 86.121% of unlabeled GFP-ADSCs (A) and about 83.052% of labeled GFP-ADSCs (B) were in proliferative state (S phase).

Abbreviations: GFP-ADSCs, green fluorescent protein adipose-derived stem cells; FCM, Flow Cytometry Method.

Figure S3 Cell cycle of labeled or unlabeled GFP-ADSCs with 25 μg/ml SPIO.Notes: FCM showed that about 86.121% of unlabeled GFP-ADSCs (A) and about 83.052% of labeled GFP-ADSCs (B) were in proliferative state (S phase).Abbreviations: GFP-ADSCs, green fluorescent protein adipose-derived stem cells; FCM, Flow Cytometry Method.

Figure S4 Prussian blue staining, TEM examination of the control groups.

Notes: (A) Prussian blue staining showed no blue-stained iron particles in the lesion area of the control animals. (B) TEM further demonstrated no iron particles accumulated in the lesion area of the control animals. Scale bar A measures 100 μm, B measures 2 μm.

Abbreviation: TEM, transmission electron microscope.

Figure S4 Prussian blue staining, TEM examination of the control groups.Notes: (A) Prussian blue staining showed no blue-stained iron particles in the lesion area of the control animals. (B) TEM further demonstrated no iron particles accumulated in the lesion area of the control animals. Scale bar A measures 100 μm, B measures 2 μm.Abbreviation: TEM, transmission electron microscope.

Figure S5 Histological Examination of the injured carotid artery at day 7 and 30 following transplantation.

Notes: (A) Macrophage mark MAC-3 could be detected at 7 d. (B) The number of macrophage was obviously decreasing at 30 d. Scale bar measures 100 μm.