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Infrared fluorescent protein 1.4 genetic labeling tracks engrafted cardiac progenitor cells in mouse ischemic hearts.

Chen L, Phillips MI, Miao HL, Zeng R, Qin G, Kim IM, Weintraub NL, Tang Y - PLoS ONE (2014)

Bottom Line: Lentiviral mediated IFP1.4 gene labeling is stable, and does not impact the apoptosis and cardiac differentiation of CPC.At 1 week after injection, 70% of the NIRF signal was lost when compared to the intensity of the day 1 signal.Our studies have shown that IFP1.4 gene labeling can be used to track the viability of transplanted cells in vivo.

View Article: PubMed Central - PubMed

Affiliation: Department of Cardiology, Zhongda Hospital, Medical School of Southeast University, Nanjing, China; Department of Medicine, University of Cincinnati, Cincinnati, Ohio, United States of America.

ABSTRACT
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.

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Assessment of CPC survival in ischemic myocardium using in vivo near infrared fluorescent (NIRF) imaging.(A–B) Representative near-infrared fluorescence imaging in mice obtained at 1d, 1w and 2w post cell transplantation (n = 6), the fluorescence image was overlaid on an X-ray using a KODAK In-Vivo Multispectral FX Image 2D Station.
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pone-0107841-g005: Assessment of CPC survival in ischemic myocardium using in vivo near infrared fluorescent (NIRF) imaging.(A–B) Representative near-infrared fluorescence imaging in mice obtained at 1d, 1w and 2w post cell transplantation (n = 6), the fluorescence image was overlaid on an X-ray using a KODAK In-Vivo Multispectral FX Image 2D Station.

Mentions: To determine whether lenti-IFP1.4 cell labeling can be used for in vivo tracking of cell survival in ischemic hearts following cell transplantation, CPCs were genetically modified by lentiviral vector with IFP1.4 (CPC IFP1.4). Mice then underwent left anterior descending artery ligation and intramyocardial injection with CPC IFP1.4. Noninvasive near infrared fluorescence and X-ray image overlays were conducted at day 1, day 7 and day 14 after cell transplantation to track the survival of transplanted cells in vivo. A strong near infrared signal was observed in mice treated with IFP1.4-labeled CPC at 1 day post cell transplantation; however, signal strength was significantly reduced by 70% 1 week post cell transplantation, and further reduced by 88% at 2 weeks post cell transplantation (Fig. 5A–B, n = 6).


Infrared fluorescent protein 1.4 genetic labeling tracks engrafted cardiac progenitor cells in mouse ischemic hearts.

Chen L, Phillips MI, Miao HL, Zeng R, Qin G, Kim IM, Weintraub NL, Tang Y - PLoS ONE (2014)

Assessment of CPC survival in ischemic myocardium using in vivo near infrared fluorescent (NIRF) imaging.(A–B) Representative near-infrared fluorescence imaging in mice obtained at 1d, 1w and 2w post cell transplantation (n = 6), the fluorescence image was overlaid on an X-ray using a KODAK In-Vivo Multispectral FX Image 2D Station.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4214633&req=5

pone-0107841-g005: Assessment of CPC survival in ischemic myocardium using in vivo near infrared fluorescent (NIRF) imaging.(A–B) Representative near-infrared fluorescence imaging in mice obtained at 1d, 1w and 2w post cell transplantation (n = 6), the fluorescence image was overlaid on an X-ray using a KODAK In-Vivo Multispectral FX Image 2D Station.
Mentions: To determine whether lenti-IFP1.4 cell labeling can be used for in vivo tracking of cell survival in ischemic hearts following cell transplantation, CPCs were genetically modified by lentiviral vector with IFP1.4 (CPC IFP1.4). Mice then underwent left anterior descending artery ligation and intramyocardial injection with CPC IFP1.4. Noninvasive near infrared fluorescence and X-ray image overlays were conducted at day 1, day 7 and day 14 after cell transplantation to track the survival of transplanted cells in vivo. A strong near infrared signal was observed in mice treated with IFP1.4-labeled CPC at 1 day post cell transplantation; however, signal strength was significantly reduced by 70% 1 week post cell transplantation, and further reduced by 88% at 2 weeks post cell transplantation (Fig. 5A–B, n = 6).

Bottom Line: Lentiviral mediated IFP1.4 gene labeling is stable, and does not impact the apoptosis and cardiac differentiation of CPC.At 1 week after injection, 70% of the NIRF signal was lost when compared to the intensity of the day 1 signal.Our studies have shown that IFP1.4 gene labeling can be used to track the viability of transplanted cells in vivo.

View Article: PubMed Central - PubMed

Affiliation: Department of Cardiology, Zhongda Hospital, Medical School of Southeast University, Nanjing, China; Department of Medicine, University of Cincinnati, Cincinnati, Ohio, United States of America.

ABSTRACT
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.

Show MeSH
Related in: MedlinePlus