Lentiviral transduction and subsequent loading with nanoparticles do not affect cell viability and proliferation in hair‐follicle‐bulge‐derived stem cells in vitro. (15th December 2016)

• Record Type: Journal Article
• Authors: Schomann, Timo; Mezzanotte, Laura; Lourens, Ierry‐Ann‐Lym M.; de Groot, John C. M. J.; Frijns, Johan H. M.; Huisman, Margriet A.
• Is Part Of: Contrast media & molecular imaging. Volume 11:Number 6(2016:Nov./Dec.)
• Page Start: 550
• Page End: 560
• Publication Date: 2016-12-15
• Subjects: stem cells -- hair follicle -- bioluminescence -- fluorescence -- magnetic resonance imaging -- magnetic nanoparticles; Diagnostic imaging -- Periodicals; Magnetic resonance imaging -- Periodicals; Contrast media (Diagnostic imaging) -- Periodicals; Contrast Media -- Periodicals; Diagnostic Imaging -- Periodicals; Substances de contraste -- Périodiques; Diagnostics moléculaires -- Périodiques; Imagerie médicale; Substance de contraste; Périodique électronique (Descripteur de forme); Ressource Internet (Descripteur de forme); 616.0754
• DOI: 10.1002/cmmi.1717
• Languages: English
• ISSNs: 1555-4309

Abstract

Abstract : The application of stem cells in the treatment of various degenerative diseases is highly promising. However, cell‐based therapy could be limited by the problem of low viability of grafted cells and uncertainty about their fate. The combination of molecular imaging and contrast‐enhanced MRI may give more insight into the survival and behavior of grafted stem cells. We explore hair‐follicle‐bulge‐derived stem cells (HFBSCs) as a potential candidate for autologous cell‐based therapy. HFBSCs are transduced with a lentiviral construct with genes coding for bioluminescent (Luc2) and fluorescent (copGFP) reporter proteins, and subsequently loaded with magnetic nanoparticles to enable MRI visualization. Thus, we investigate for the first time if lentiviral transduction and cellular loading with nanoparticles have a cytotoxic effect upon these stem cells. Transduction efficiency, proliferation rate, cell viability and reporter protein co‐expression during long‐term culture of transduced HFBSCs were studied using fluorescence and bioluminescence microscopy. In addition, the effect of TMSR50 nanoparticles on proliferation and viability was investigated using the MTS assay and bioluminescence microscopy. The amount of TMSR50‐loaded HFBSCs needed to reach signal threshold for MRI was assessed using an agarose phantom. Transduction with the Luc2‐copGFP construct did not influence senescence, proliferation, doubling time, and differentiation of the HFBSCs. CopGFP expression was visible immediately after transduction and persisted for at least 15 passages, concomitantly with Luc2 expression. Cellular loading with TMSR50 nanoparticles did not affect cell viability and proliferation. The results imply that combined MRI and bioluminescence imaging may enable in vivo localization and long‐term monitoring of grafted viable HFBSCs. Copyright © 2016 John Wiley & Sons, Ltd. Abstract : Transduction of hair‐follicle‐bulge‐derived stem cells with a Luc2‐copGFP construct followed by subsequent loading with red‐fluorescent iron‐containing TMSR50 nanoparticles results in equimolar expression of both reporter molecules, it does not interfere with cellular senescence, cell viability, proliferation and differentiation and enables in vitro detection by means of fluorescence imaging (IVIS) and MRI. These results imply that MRI and molecular optical imaging can be combined to enable in vivo localization and long‐term monitoring of viable hair‐follicle‐bulge‐derived stem cells after engraftment.