AAV-iRFP labelling of human mesenchymal stem cells for near-infrared fluorescence imaging.

Can Huang,Qin Chen,Wenjun Lan,Xiaomei Liu,Feifei Wang,Chun Zhang

doi:10.1042/bsr20160556

Abstract

Near-IR fluorescence (NIRF) imaging is a new technology using IR fluorescent protein (iRFP) gene labelling and is potentially useful for in vivo applications. In the present study, we expressed iRFP and the TNF-related apoptosis inducing ligand gene in mesenchymal stem cells (MSCs) using adeno-associated virus (AAV) and showed that iRFP-labelled MSCs can be detected by fluorescence microscopy. We injected mice with MSCs labelled with AAV-iRFP, which we were then able to detect by whole-animal NIRF imaging. Our technique provides a visualizable, convenient and sensitive platform for research on tracking the fate of transplanted MSC cells in vivo.

Highlights

Human mesenchymal stem cells (MSCs) are multipotent cells that can self-renew, proliferate and differentiate into various cell types
IRFP682 was linked to the previous plasmid vector digested by SalI/BamHI to obtain a recombinant vector with the iRFP682 gene (5292 bp) (Figure 2 E)
CMV-promoter was linked to the previous plasmid vector digested by BglII to obtain recombinant vector adeno-associated virus plasmid vector (pAAV)-iRFP682-multiple clone site (MCS) with two complete expression cassettes (6541 bp)

Summary

Introduction

Human mesenchymal stem cells (MSCs) are multipotent cells that can self-renew, proliferate and differentiate into various cell types. They can overcome difficulties associated with immune rejection of transplanted cells and can be isolated from the body, cultured in vitro and transplanted into patients autologously. They have a high metabolic activity and robustly express transgenes. The properties of MSCs may have an important role in protecting cells from immune responses against the transgene that they are carrying [4]. In order to use MSCs as transgene delivery vehicles, a high percentage of the cells need to become infected in order to express high levels of transgene and this process must be safe

Methods

Results

Conclusion