Magnetic Resonance Imaging of Human Embryonic Stem Cells

Jaehoon Chung1, Mayumi Yamada1, Phillip C. Yang1

1 Stanford University School of Medicine, Stanford, California
Publication Name:  Current Protocols in Stem Cell Biology
Unit Number:  Unit 5A.3
DOI:  10.1002/9780470151808.sc05a03s10
Online Posting Date:  August, 2009
GO TO THE FULL TEXT: PDF or HTML at Wiley Online Library

Abstract

Magnetic resonance imaging (MRI) may emerge as an ideal non‐invasive imaging modality to monitor stem cell therapy in the failing heart. This imaging modality generates any arbitrary tomographic view at high spatial and temporal resolution with exquisite intrinsic tissue contrast. This capability enables robust evaluation of both the cardiac anatomy and function. Traditionally, superparamagnetic iron oxide nanoparticle (SPIO) has been widely used for cellular MRI due to SPIO's ability to enhance sensitivity of MRI by inducing remarkable hypointense, negative signal, “blooming effect” on T2*‐weighted MRI acquisition. Recently, manganese chloride (MnCl2) has been reported by our laboratory for its ability as a contrast agent to track biological activity of viable cells. Hyperintense, positive signals can be achieved from the Mn2+‐labeled stem cells on T1‐weighted MRI acquisition. Cytotoxicity is a potential drawback of Mn2+ labeling of the cells. However, in our laboratory the labeling method has been optimized to minimize cytotoxic effects. This article describes two different magnetic labeling methods of human embryonic stem cells (hESC) using SPIO and MnCl2. Curr. Protoc. Stem Cell Biol. 10:5A.3.1‐5A.3.9. © 2009 by John Wiley & Sons, Inc.

Keywords: human embryonic stem cell (hESC); magnetic resonance imaging (MRI); superparamagnetic iron oxide (SPIO); manganese

     
 
GO TO THE FULL PROTOCOL:
PDF or HTML at Wiley Online Library

Table of Contents

  • Introduction
  • Basic Protocol 1: Direct Magnetic Labeling of Human Embryonic Stem Cells (hESC) Using SPIO
  • Alternate Protocol 1: Direct Labeling of Human Embryonic Stem Cells Using Manganese Chloride
  • Reagents and Solutions
  • Commentary
  • Literature Cited
  • Figures
     
 
GO TO THE FULL PROTOCOL:
PDF or HTML at Wiley Online Library

Materials

Basic Protocol 1: Direct Magnetic Labeling of Human Embryonic Stem Cells (hESC) Using SPIO

  Materials
  • H9 hESC (WiCell)
  • hESC medium (see recipe)
  • SPIO (Feridex, Bayer Healthcare Pharmaceuticals)
  • Protamine sulfate (PS, American Pharmaceutical Partners)
  • Phosphate‐buffered saline, calcium‐ and magnesium‐free (CMF‐PBS; see recipe)
  • Heparin sodium (Fujisawa)
  • Phantom (see recipe)
  • Matrigel‐coated 100‐mm tissue culture dishes (BD Biosciences; coat dishes according to manufacturer's instructions)
  • 0.2‐ml PCR tubes
  • Signa 3T Excite HD scanner (GE Health System) and an array knee coil with GRE or SPGR sequence

Alternate Protocol 1: Direct Labeling of Human Embryonic Stem Cells Using Manganese Chloride

  Materials
  • H9 hESC (WiCell)
  • hESC medium (see recipe)
  • TrypLE express (Invitrogen)
  • MnCl 2 (Sigma)
  • 0.9% (w/v) sodium chloride (9 mg sodium chloride/ml of distilled water)
  • Phantom (see recipe)
  • Matrigel‐coated 100‐mm tissue culture dishes (BD Biosciences; coat dishes according to manufacturer's instructions)
  • Hemacytometer
  • 15‐ml conical tubes
  • 0.2‐ml PCR tubes
  • Signa 3T Excite HD scanner (GE Health System)
GO TO THE FULL PROTOCOL:
PDF or HTML at Wiley Online Library

Figures

Videos

Literature Cited

Literature Cited
   Aoki, I., Takahashi, Y., Chuang, K.H., Silva, A.C., Igarashi, T., Tanaka, C., Childs, R.W., and Korefsky, A.P. 2006. Cell labeling for magnetic resonance imaging with the T1 agent manganese chloride. NMR Biomed. 19:50‐59.
   Auerbach, M.A., Schoder, H., Hoh, C., Gambhir, S.S., Yaghoubi, S., Sayre, J.B., Silverman, D., Phelps, M.E., Schelbert, H.R., and Czernin, J. 1999. Prevalence of myocardial viability as detected by positron emission tomography in patients with ischemic cardiomyopathy. Circulation 99:2921‐2926.
   Bruvold, M., Nordhoy, W., Anthonsen, H.W., Brurok, H., and Jynge, P. 2005. Manganese‐calcium interactions with contrast media for cardiac magnetic resonance imaging: A study of manganese chloride supplemented with calcium gluconate in isolated Guinea pig hearts. Invest. Radiol. 40:117‐125.
   Bulte, J.W., Douglas, T., Witwer, B., Zhang, S.C., Strable, E., Lewis, B.K., Zwicke, H., Miller, B., van Geleren, P., Moscovitz, B.M., Duncan, I.D., and Frank, J.A. 2001. Magnetodendrimers allow endosomal magnetic labeling and in vivo tracking of stem cells. Nat. Biotechnol. 19:1141‐1147.
   Chen, I.Y., Greve, J.M., Gheysens, O., Willmann, J.K., Rodriguez‐Porcel, M., Chu, P., Sheikh, A.Y., Faranesh, A.Z., Paulmurugen, R., Yang, P.C., Wu, J.C., and Gambhir, S.S. 2008. Comparison of optical bioluminescence reporter gene and superparamagnetic iron oxide MR contrast agent as cell markers for noninvasive imaging of cardiac cell transplantation. Mol. Imaging Biol. 11:178‐187.
   Frank, J.A., Zywicke, H., Jordan, E.K., Mitchell, J., Lewis, B.K., Miller, B., Bryant, L.H. Jr., and Bulte, J.W. 2002. Magnetic intracellular labeling of mammalian cells by combining (FDA‐approved) superaramagnetic iron oxide MR contrast agents and commonly used transfection agents. Acad. Radiol. 9:S484‐S487.
   Kostura, L., Kraitchman, D.L., Mackay, E.M., Pittinger, M.F., and Bulte, J.W. 2004. Feridex labeling of mesenchymal stem cells inhibits chondrogenesis but not adipogenesis or osteogenesis. NMR Biomed. 17:513‐517.
   Kraitchman, D.L., Heldman, A.W., Atalar, E., Amado, L.C., Martin, B.J., Pittenger, M.F., Hare, J.M., and Bulte, J.W. 2003. In vivo magnetic resonance imaging of mesenchymal stem cells in myocardial infarction. Circulation 107:2290‐2293.
   Li, Z., Suzuki, Y., Huang, M., Cao, F., Xie, X., Connolly, A.J., Yang, P.C., and Wu, J.C. 2008. Comparison of reporter gene and iron particle labeling for tracking fate of human embryonic stem cells and differentiated endothelial cells in living subjects. Stem Cells 26:864‐873.
   Lin, Y.J. and Koretsky, A.P. 1997. Manganese ion enhances T1‐weighted MRI during brain activation: An approach to direct imaging of brain function. Magn. Reson. Med. 38:378‐388.
   Suzuki, Y., Zhang, S., Kundu, P., Yeung, A.C., Robbins, R.C., and Yang, P.C. 2007. In vitro comparison of the biological effects of three transfection methods for magnetically labeling mouse embryonic stem cells with ferumoxides. Magn. Reson. Med. 57:1173‐1179.
   Walczak, P., Kedziorek, D.A., Gilad, A.A., Lin, S., and Bulte, J.W. 2005. Instant MR labeling of stem cells using magnetoelectroporation. Magn. Reson. Med. 54:769‐774.
   Wu, J.C., Sundaresan, G., Iyer, M., and Gambhir, S.S. 2001. Noninvasive optical imaging of firefly luciferase reporter gene expression in skeletal muscles of living mice. Mol. Ther. 4:297‐306.
   Wu, J.C., Chen, I.Y., Sundaresan, G., Min, J.J., De, A., Qiao, J.H., Fishbein, M.C., and Gambhir, S.S. 2003. Molecular imaging of cardiac cell transplantation in living animals using optical bioluminescence and positron emission tomography. Circulation 108:1302‐1305.
GO TO THE FULL PROTOCOL:
PDF or HTML at Wiley Online Library