Assessing in vitro stem‐cell function and tracking engraftment of stem cells in ischaemic hearts by using novel iRFP gene labelling

Journal of Cellular and Molecular Medicine

Yingjie Wang²  Mi Zhou³  Xiaolong Wang²  Gangjian Qin¹  Neal L. Weintraub³ 

[1] Feinberg Cardiovascular Research Institute, Northwestern University Feinberg School of Medicine, Chicago, IL, USA;Internal Medicine of Traditional Chinese Medicine, Shuguang Hospital of Shanghai University of Traditional Chinese Medicine, Shanghai, China;Department of Medicine, University of Cincinnati, Cincinnati, OH, USA

关键词: stem cells;    myocardial infarction;    cell transplantation;    iRFP;    GFP;   

DOI  :  10.1111/jcmm.12321

来源: Wiley

【 摘 要 】

Abstract

Near-infrared fluorescence (NIRF) imaging by using infrared fluorescent protein (iRFP) gene labelling is a novel technology with potential value for in vivo applications. In this study, we expressed iRFP in mouse cardiac progenitor cells (CPC) by lentiviral vector and demonstrated that the iRFP-labelled CPC (CPC^iRFP) can be detected by flow cytometry and fluorescent microscopy. We observed a linear correlation in vitro between cell numbers and infrared signal intensity by using the multiSpectral imaging system. CPC^iRFP injected into the non-ischaemic mouse hindlimb were also readily detected by whole-animal NIRF imaging. We then compared iRFP against green fluorescent protein (GFP) for tracking survival of engrafted CPC in mouse ischaemic heart tissue. GFP-labelled CPC (CPC^GFP) or CPC labelled with both iRFP and GFP (CPC^{iRFP GFP}) were injected intramyocardially into mouse hearts after infarction. Three days after cell transplantation, a strong NIRF signal was detected in hearts into which CPC^{iRFP GFP}, but not CPC^GFP, were transplanted. Furthermore, iRFP fluorescence from engrafted CPC^{iRFP GFP} was detected in tissue sections by confocal microscopy. In conclusion, the iRFP-labelling system provides a valuable molecular imaging tool to track the fate of transplanted progenitor cells in vivo.

【 授权许可】

CC BY   
© 2014 The Authors. Journal of Cellular and Molecular Medicine published by John Wiley & Sons Ltd and Foundation for Cellular and Molecular Medicine.

Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.