Imaging and Tracking Stem Cell Engraftment in Ischemic Hearts by Near-Infrared Fluorescent Protein (iRFP) Labeling

  • Xuan Su
  • Yan Shen
  • Neal L. Weintraub
  • Yaoliang TangEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 2150)


Stem cell-based therapies hold great promise as alternative therapeutic strategies for various chronic diseases, including ischemic cardiomyopathy. Tracking the engraftment of transplanted stem cells is critical to the assessment of donor cell survival in the host environment. Fluorescent proteins, such as green fluorescent protein (GFP), have been widely used to track the fate of donor cells; however, GFP labeling has limitations with regard to noninvasively measuring cell engraftment in vivo. Our research indicates that near-infrared fluorescent protein (iRFP) labeling offers advantages for noninvasive in vivo imaging and histological assessment. Here, we present a protocol for using the lentiviral vector-mediated iRFP vector system to label and track donor stem cells in ischemic mouse hearts.


Stem cells iRFP Green fluorescent protein (GFP) Lentivirus Myocardial infarction 



Weintraub and Y. Tang were partially supported by the American Heart Association: GRNT31430008, NIH-AR070029, NIH-HL086555, and NIH-HL134354. All authors declare that he/she has no conflict of interest. This article does not contain any studies with human participants performed by any of the authors.


  1. 1.
    Laslett LJ, Alagona P Jr, Clark BA 3rd, Drozda JP Jr, Saldivar F, Wilson SR, Poe C, Hart M (2012) The worldwide environment of cardiovascular disease: prevalence, diagnosis, therapy, and policy issues: a report from the American College of Cardiology. J Am Coll Cardiol 60:S1–S49CrossRefGoogle Scholar
  2. 2.
    Ju C, Li Y, Shen Y, Liu Y, Cai J, Liu N, Ma G, Tang Y (2018) Transplantation of cardiac mesenchymal stem cell-derived exosomes for angiogenesis. J Cardiovasc Transl Res 11:429–437CrossRefGoogle Scholar
  3. 3.
    Ju C, Shen Y, Ma G, Liu Y, Cai J, Kim IM, Weintraub NL, Liu N, Tang Y (2018) Transplantation of cardiac mesenchymal stem cell-derived exosomes promotes repair in ischemic myocardium. J Cardiovasc Transl Res 11:420–428CrossRefGoogle Scholar
  4. 4.
    Su X, Jin Y, Shen Y, Ju C, Cai J, Liu Y, Kim IM, Wang Y, Yu H, Weintraub NL, Jiang M, Tang Y (2018) Exosome-derived dystrophin from allograft myogenic progenitors improves cardiac function in duchenne muscular dystrophic mice. J Cardiovasc Transl Res 11:412–419CrossRefGoogle Scholar
  5. 5.
    Wang Z, Su X, Ashraf M, Kim IM, Weintraub NL, Jiang M, Tang Y (2018) Regenerative therapy for cardiomyopathies. J Cardiovasc Transl Res 11:357–365CrossRefGoogle Scholar
  6. 6.
    Chalfie M, Tu Y, Euskirchen G, Ward WW, Prasher DC (1994) Green fluorescent protein as a marker for gene expression. Science 263:802–805CrossRefGoogle Scholar
  7. 7.
    Frangioni JV (2003) In vivo near-infrared fluorescence imaging. Curr Opin Chem Biol 7:626–634CrossRefGoogle Scholar
  8. 8.
    Wang Y, Zhang L, Pan Y, Chen L, Weintraub N, Tang Y (2013) Identification of stem cells after transplantation. Methods Mol Biol 1036:89–94CrossRefGoogle Scholar
  9. 9.
    Shaner NC, Steinbach PA, Tsien RY (2005) A guide to choosing fluorescent proteins. Nat Methods 2:905–909CrossRefGoogle Scholar
  10. 10.
    Lecoq J, Schnitzer MJ (2011) An infrared fluorescent protein for deeper imaging. Nat Biotechnol 29:715–716CrossRefGoogle Scholar
  11. 11.
    Chen G, Zhang Y, Li C, Huang D, Wang Q, Wang Q (2018) Recent advances in tracking the transplanted stem cells using near-infrared fluorescent nanoprobes: turning from the first to the second near-infrared window. Adv Healthc Mater 7:e1800497CrossRefGoogle Scholar
  12. 12.
    Wang Y, Zhou M, Wang X, Qin G, Weintraub NL, Tang Y (2014) Assessing in vitro stem-cell function and tracking engraftment of stem cells in ischaemic hearts by using novel iRFP gene labelling. J Cell Mol Med 18:1889–1894CrossRefGoogle Scholar
  13. 13.
    Chen L, Phillips MI, Miao HL, Zeng R, Qin G, Kim IM, Weintraub NL, Tang Y (2014) Infrared fluorescent protein 1.4 genetic labeling tracks engrafted cardiac progenitor cells in mouse ischemic hearts. PLoS One 9:e107841CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2019

Authors and Affiliations

  • Xuan Su
    • 1
  • Yan Shen
    • 1
  • Neal L. Weintraub
    • 1
  • Yaoliang Tang
    • 1
    Email author
  1. 1.Vascular Biology Center, Medical College of GeorgiaAugusta UniversityAugustaUSA

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