Infrared Fluorescent Protein 1.4 Genetic Labeling Tracks Engrafted Cardiac Progenitor Cells in Mouse Ischemic Hearts

Stem cell therapy has a potential for regenerating damaged myocardium. However, a key obstacle to cell therapy’s success is the loss of engrafted cells due to apoptosis or necrosis in the ischemic myocardium. While many strategies have been developed to improve engrafted cell survival, tools to eval...

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Main Authors:	Chen, Lijuan, Phillips, M. Ian, Miao, Hui-Lai, Zeng, Rong, Qin, Gangjian, Kim, Il-man, Weintraub, Neal L., Tang, Yaoliang
Format:	Online
Language:	English
Published:	Public Library of Science 2014
Online Access:	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4214633/

id	pubmed-4214633
recordtype	oai_dc
spelling	pubmed-42146332014-11-05 Infrared Fluorescent Protein 1.4 Genetic Labeling Tracks Engrafted Cardiac Progenitor Cells in Mouse Ischemic Hearts Chen, Lijuan Phillips, M. Ian Miao, Hui-Lai Zeng, Rong Qin, Gangjian Kim, Il-man Weintraub, Neal L. Tang, Yaoliang Research Article Stem cell therapy has a potential for regenerating damaged myocardium. However, a key obstacle to cell therapy’s success is the loss of engrafted cells due to apoptosis or necrosis in the ischemic myocardium. While many strategies have been developed to improve engrafted cell survival, tools to evaluate cell efficacy within the body are limited. Traditional genetic labeling tools, such as GFP-like fluorescent proteins (eGFP, DsRed, mCherry), have limited penetration depths in vivo due to tissue scattering and absorption. To circumvent these limitations, a near-infrared fluorescent mutant of the DrBphP bacteriophytochrome from Deinococcus radiodurans, IFP1.4, was developed for in vivo imaging, but it has yet to be used for in vivo stem/progenitor cell tracking. In this study, we incorporated IFP1.4 into mouse cardiac progenitor cells (CPCs) by a lentiviral vector. Live IFP1.4-labeled CPCs were imaged by their near-infrared fluorescence (NIRF) using an Odyssey scanner following overnight incubation with biliverdin. A significant linear correlation was observed between the amount of cells and NIRF signal intensity in in vitro studies. Lentiviral mediated IFP1.4 gene labeling is stable, and does not impact the apoptosis and cardiac differentiation of CPC. To assess efficacy of our model for engrafted cells in vivo, IFP1.4-labeled CPCs were intramyocardially injected into infarcted hearts. NIRF signals were collected at 1-day, 7-days, and 14-days post-injection using the Kodak in vivo multispectral imaging system. Strong NIRF signals from engrafted cells were imaged 1 day after injection. At 1 week after injection, 70% of the NIRF signal was lost when compared to the intensity of the day 1 signal. The data collected 2 weeks following transplantation showed an 88% decrease when compared to day 1. Our studies have shown that IFP1.4 gene labeling can be used to track the viability of transplanted cells in vivo. Public Library of Science 2014-10-30 /pmc/articles/PMC4214633/ /pubmed/25357000 http://dx.doi.org/10.1371/journal.pone.0107841 Text en © 2014 Chen et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
repository_type	Open Access Journal
institution_category	Foreign Institution
institution	US National Center for Biotechnology Information
building	NCBI PubMed
collection	Online Access
language	English
format	Online
author	Chen, Lijuan Phillips, M. Ian Miao, Hui-Lai Zeng, Rong Qin, Gangjian Kim, Il-man Weintraub, Neal L. Tang, Yaoliang
spellingShingle	Chen, Lijuan Phillips, M. Ian Miao, Hui-Lai Zeng, Rong Qin, Gangjian Kim, Il-man Weintraub, Neal L. Tang, Yaoliang Infrared Fluorescent Protein 1.4 Genetic Labeling Tracks Engrafted Cardiac Progenitor Cells in Mouse Ischemic Hearts
author_facet	Chen, Lijuan Phillips, M. Ian Miao, Hui-Lai Zeng, Rong Qin, Gangjian Kim, Il-man Weintraub, Neal L. Tang, Yaoliang
author_sort	Chen, Lijuan
title	Infrared Fluorescent Protein 1.4 Genetic Labeling Tracks Engrafted Cardiac Progenitor Cells in Mouse Ischemic Hearts
title_short	Infrared Fluorescent Protein 1.4 Genetic Labeling Tracks Engrafted Cardiac Progenitor Cells in Mouse Ischemic Hearts
title_full	Infrared Fluorescent Protein 1.4 Genetic Labeling Tracks Engrafted Cardiac Progenitor Cells in Mouse Ischemic Hearts
title_fullStr	Infrared Fluorescent Protein 1.4 Genetic Labeling Tracks Engrafted Cardiac Progenitor Cells in Mouse Ischemic Hearts
title_full_unstemmed	Infrared Fluorescent Protein 1.4 Genetic Labeling Tracks Engrafted Cardiac Progenitor Cells in Mouse Ischemic Hearts
title_sort	infrared fluorescent protein 1.4 genetic labeling tracks engrafted cardiac progenitor cells in mouse ischemic hearts
description	Stem cell therapy has a potential for regenerating damaged myocardium. However, a key obstacle to cell therapy’s success is the loss of engrafted cells due to apoptosis or necrosis in the ischemic myocardium. While many strategies have been developed to improve engrafted cell survival, tools to evaluate cell efficacy within the body are limited. Traditional genetic labeling tools, such as GFP-like fluorescent proteins (eGFP, DsRed, mCherry), have limited penetration depths in vivo due to tissue scattering and absorption. To circumvent these limitations, a near-infrared fluorescent mutant of the DrBphP bacteriophytochrome from Deinococcus radiodurans, IFP1.4, was developed for in vivo imaging, but it has yet to be used for in vivo stem/progenitor cell tracking. In this study, we incorporated IFP1.4 into mouse cardiac progenitor cells (CPCs) by a lentiviral vector. Live IFP1.4-labeled CPCs were imaged by their near-infrared fluorescence (NIRF) using an Odyssey scanner following overnight incubation with biliverdin. A significant linear correlation was observed between the amount of cells and NIRF signal intensity in in vitro studies. Lentiviral mediated IFP1.4 gene labeling is stable, and does not impact the apoptosis and cardiac differentiation of CPC. To assess efficacy of our model for engrafted cells in vivo, IFP1.4-labeled CPCs were intramyocardially injected into infarcted hearts. NIRF signals were collected at 1-day, 7-days, and 14-days post-injection using the Kodak in vivo multispectral imaging system. Strong NIRF signals from engrafted cells were imaged 1 day after injection. At 1 week after injection, 70% of the NIRF signal was lost when compared to the intensity of the day 1 signal. The data collected 2 weeks following transplantation showed an 88% decrease when compared to day 1. Our studies have shown that IFP1.4 gene labeling can be used to track the viability of transplanted cells in vivo.
publisher	Public Library of Science
publishDate	2014
url	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4214633/
_version_	1613150524754886656

Infrared Fluorescent Protein 1.4 Genetic Labeling Tracks Engrafted Cardiac Progenitor Cells in Mouse Ischemic Hearts

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