Labeling human embryonic stem-cell-derived cardiomyocytes for tracking with MR imaging
- 262 Downloads
Human embryonic stem cells (hESC) can generate cardiomyocytes (CM), which offer promising treatments for cardiomyopathies in children. However, challenges for clinical translation result from loss of transplanted cell from target sites and high cell death. An imaging technique that noninvasively and repetitively monitors transplanted hESC-CM could guide improvements in transplantation techniques and advance therapies.
To develop a clinically applicable labeling technique for hESC-CM with FDA-approved superparamagnetic iron oxide nanoparticles (SPIO) by examining labeling before and after CM differentiation.
Materials and methods
Triplicates of hESC were labeled by simple incubation with 50 μg/ml of ferumoxides before or after differentiation into CM, then imaged on a 7T MR scanner using a T2-weighted multi-echo spin-echo sequence. Viability, iron uptake and T2-relaxation times were compared between groups using t-tests.
hESC-CM labeled before differentiation demonstrated significant MR effects, iron uptake and preserved function. hESC-CM labeled after differentiation showed no significant iron uptake or change in MR signal (P < 0.05). Morphology, differentiation and viability were consistent between experimental groups.
hESC-CM should be labeled prior to CM differentiation to achieve a significant MR signal. This technique permits monitoring delivery and engraftment of hESC-CM for potential advancements of stem cell-based therapies in the reconstitution of damaged myocardium.
KeywordsHuman embryonic stem cells Cardiomyocytes Iron oxides SPIO MR imaging Cell labeling Children
This study was supported by a research grant from the California Institute for Regenerative Medicine (CIRM), grant # RS1-00381-1.
Successfully labeled, terminally differentiated, beating hESC-CM labeled with SPIO, ferumoxides, prior to CM differentiation. SPIO particles are seen in the cytoplasm of the labeled hESC-CM (MP4 13085 kb)
Terminally differentiated, beating hESC-CM that underwent attempted labeling following CM differentiation. No SPIO particles are visible in these hESC-CM (MP4 4698 kb)
Control, unlabeled, terminally differentiated, beating hESC-CM. No SPIO particles are present. These cells show no morphologic or functional difference compared to labeled cells in (1) (MP4 4072 kb)
- 12.Li SH, Lai TY, Sun Z et al (2009) Tracking cardiac engraftment and distribution of implanted bone marrow cells: comparing intra-aortic, intravenous, and intramyocardial delivery. J Thorac Cardiovasc Surg 137(1225–1233):e1221Google Scholar
- 29.Chung J, Yamada M, Yang PC (2009) Magnetic resonance imaging of human embryonic stem cells. Curr Protoc Stem Cell Biol Chapter 5:Unit 5A 3Google Scholar
- 37.Henning TD, Boddington S, Daldrup-Link HE (2008) Labeling hESCs and hMSCs with iron oxide nanoparticles for non-invasive in vivo tracking with MR imaging. J Vis Exp 31:pii 685Google Scholar