Simultaneous gene delivery and tracking of cells using fluorescent upconversion nanoparticles for cell therapy
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Simultaneous gene delivery and tracking of cells using fluorescent upconversion nanoparticles for cell therapy Muthu Kumara Gnanasammandhan Jayakumar1, Yong Zhang1,2,* 1
Division of Bioengineering, Faculty of Engineering, National University of Singapore,
117576, Singapore 2
Nanoscience and Nanotechnology Initiative, National University of Singapore,
117576, Singapore
ABSTRACT Cell therapy has attained new heights with the current developments in the field of stem cell therapy and is often done in combination with gene therapy. Much research has gone into developing safe vectors to deliver genes to the cells used for cell therapy and also in tracking the transplanted cells and determining their fate. Even though viral vectors are very efficient in gene delivery, there are major safety concerns related to it. Herein, we report the use of novel fluorescent upconversion nanoparticles (UCNs) for simultaneous gene delivery and tracking of cells for cell-based therapies. It is a safe alternative to viral vectors and offers imaging capabilities too, which will give an insight on the fate of transplanted cells.
INTRODUCTION Cell therapy has been in practice for decades with little success. But it has reached a peak in the past decade with the recent advances in stem cell-based therapies. It has become a hot field with a lot of focus given to it owing to the potential of stem cells for treating various diseases and disorders. Although many stem cell therapies using different types of stem cells like umbilical cord stem cells, hematopoietic stem cells and adult stem cells have started being used for treating different diseases like leukemia, stroke, immune deficiencies and heart diseases, this field is still in its infancy. Several more diseases have been targeted using stem cells and are in the research stage 1-3. These methods have been proved to be effective but its molecular basis, mechanism of action and the fate of these cells after transplantation is not very clear till date. Also the transfection of these cells with a therapeutic gene before transplantation or transfection of normal cells to convert them to induced pluripotent stem cells is facing some difficulties with regard to the immunogenicity and toxicity issues concerned with viral vectors. Here, we report the use of lanthanide-based fluorescent upconversion nanoparticles for simultaneous delivery of the required gene to the cells and track them by the fluorescence emitted by these nanoparticles. Since, the upconversion nanoparticles are excited by Near Infrared Light (NIR) and can be made to emit in the UV, Visible and NIR region. Since, NIR light is used for the excitation of these nanoparticles, it offers various advantages like greater tissue penetration depth, high signal to
noise ratio, low photodamage making both in-vitro and in-vivo high resolution fluorescence imaging possible 4, 5
METHODS Monodisperse mesoporous silica coated NaYF4 upconversion nanoparticles co-doped with Yb and Er was synthesized using a protocol previously reported 6. Basic
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