Author + information
- Received May 11, 2011
- Revision received July 25, 2011
- Accepted July 28, 2011
- Published online January 1, 2012.
- Koen E.A. van der Bogt, MD, PhD⁎,†,⁎ (, )
- Alwine A. Hellingman, MD, PhD†,
- Maarten A. Lijkwan, MD⁎,†,
- Ernst-Jan Bos, MD⁎,†,
- Margreet R. de Vries, BSc‡,
- Juliaan R.M. van Rappard, MS⁎,
- Michael P. Fischbein, MD⁎,
- Paul H. Quax, PhD†,‡,
- Robert C. Robbins, MD⁎,
- Jaap F. Hamming, MD, PhD† and
- Joseph C. Wu, MD, PhD§∥,⁎ ()
- ↵⁎Reprint requests and correspondence:
Dr. Koen E. A. van der Bogt, Leiden University Medical Center, K6-41, P.O. Box 9600, 2300 RC Leiden, the Netherlands. OR Dr. Joseph C. Wu, Stanford University School of Medicine, 265 Campus Drive, G1120B, Stanford, California 94305-5454
Objectives This study aims to provide insight into cellular kinetics using molecular imaging after different transplantation methods of bone marrow–derived mononuclear cells (MNCs) in a mouse model of peripheral artery disease (PAD).
Background MNC therapy is a promising treatment for PAD. Although clinical translation has already been established, there is a lack of knowledge about cell behavior after transplantation and about the mechanism whereby MNC therapy might ameliorate complaints of PAD.
Methods MNCs were isolated from F6 transgenic mice (FVB background) that express firefly luciferase (Fluc) and green fluorescence protein (GFP). Male FVB and C57Bl6 mice (n = 50) underwent femoral artery ligation and were randomized into 4 groups receiving the following: 1) single intramuscular (IM) injection of 2 × 106 MNCs; 2) 4 weekly IM injections of 5 × 105 MNCs; 3) 2 × 106 MNCs intravenously; and 4) phosphate-buffered saline as control. Cells were characterized by flow cytometry and in vitro bioluminescence imaging (BLI). Cell survival, proliferation, and migration were monitored by in vivo BLI, which was validated by ex vivo BLI, post-mortem immunohistochemistry, and flow cytometry. Paw perfusion and neovascularization was measured with laser Doppler perfusion imaging (LDPI) and histology, respectively.
Results In vivo BLI revealed near-complete donor cell death 4 weeks after IM transplantation. After intravenous transplantation, BLI revealed that cells migrated to the injured area in the limb, as well as to the liver, spleen, and bone marrow. Ex vivo BLI showed presence of MNCs in the scar tissue and adductor muscle. However, no significant effects on neovascularization were observed, as monitored by LDPI and histology.
Conclusions This is one of the first studies to assess kinetics of transplanted MNCs in PAD using in vivo molecular imaging. MNC survival is short-lived, MNCs do not preferentially home to injured areas, and MNCs do not significantly stimulate perfusion in this particular model.
Dr. van der Bogt is supported by the Michaël van Vloten fund. Dr. Hellingman is supported by the TeRM Smart Mix Program of the Netherlands Ministry of Economic Affairs and the Netherlands Ministry of Education, Culture and Science. Dr. Robbins is supported by National Institutes of Health Grant No. U01HL099776. Dr. Wu is supported by the Burroughs Wellcome Foundation Career Award for Medical Scientists and National Institutes of Health Grants No. RC1HL099117, R01HL093172, and R01EB009689. All authors have reported that they have no relationships relevant to the contents of this paper to disclose. Drs. van der Bogt and Hellingman contributed equally to this study.
- Received May 11, 2011.
- Revision received July 25, 2011.
- Accepted July 28, 2011.
- American College of Cardiology Foundation