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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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Analysis of CPC apoptosis and cardiac differentiation capacity after IFP1.4 gene modification.(A–B) Comparison of cell apoptosis between un-infected CPC and IFP1.4 infected CPC by TUNEL staining; (C) Real time RT-PCR comparison of cTnI expression between un-infected CPC and IFP1.4 infected CPC with/without TSA treatment.
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pone-0107841-g004: Analysis of CPC apoptosis and cardiac differentiation capacity after IFP1.4 gene modification.(A–B) Comparison of cell apoptosis between un-infected CPC and IFP1.4 infected CPC by TUNEL staining; (C) Real time RT-PCR comparison of cTnI expression between un-infected CPC and IFP1.4 infected CPC with/without TSA treatment.

Mentions: To determine whether genetic labeling with IFP 1.4 impacts on cell survival, we compare the apoptotic cells between un-infected and IFP1.4 infected CPC by TUNEL assay, there is no difference in cell apoptosis between two groups of cells (Fig. 4A–B, P>0.05, n = 7).


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)

Analysis of CPC apoptosis and cardiac differentiation capacity after IFP1.4 gene modification.(A–B) Comparison of cell apoptosis between un-infected CPC and IFP1.4 infected CPC by TUNEL staining; (C) Real time RT-PCR comparison of cTnI expression between un-infected CPC and IFP1.4 infected CPC with/without TSA treatment.
© Copyright Policy
Related In: Results  -  Collection

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

pone-0107841-g004: Analysis of CPC apoptosis and cardiac differentiation capacity after IFP1.4 gene modification.(A–B) Comparison of cell apoptosis between un-infected CPC and IFP1.4 infected CPC by TUNEL staining; (C) Real time RT-PCR comparison of cTnI expression between un-infected CPC and IFP1.4 infected CPC with/without TSA treatment.
Mentions: To determine whether genetic labeling with IFP 1.4 impacts on cell survival, we compare the apoptotic cells between un-infected and IFP1.4 infected CPC by TUNEL assay, there is no difference in cell apoptosis between two groups of cells (Fig. 4A–B, P>0.05, n = 7).

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