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  • Micro-Computed Tomography Detection of Gold Nanoparticle-Labelled Mesenchymal Stem Cells in the Rat Subretinal Layer.

Micro-Computed Tomography Detection of Gold Nanoparticle-Labelled Mesenchymal Stem Cells in the Rat Subretinal Layer.

International journal of molecular sciences (2017-02-18)
Pooi Ling Mok, Sue Ngein Leow, Avin Ee-Hwan Koh, Hairul Harun Mohd Nizam, Suet Lee Shirley Ding, Chi Luu, Raduan Ruhaslizan, Hon Seng Wong, Wan Haslina Wan Abdul Halim, Min Hwei Ng, Ruszymah Binti Hj Idrus, Shiplu Roy Chowdhury, Catherine Mae-Lynn Bastion, Suresh Kumar Subbiah, Akon Higuchi, Abdullah A Alarfaj, Kong Yong Then
ABSTRACT

Mesenchymal stem cells are widely used in many pre-clinical and clinical settings. Despite advances in molecular technology; the migration and homing activities of these cells in in vivo systems are not well understood. Labelling mesenchymal stem cells with gold nanoparticles has no cytotoxic effect and may offer suitable indications for stem cell tracking. Here, we report a simple protocol to label mesenchymal stem cells using 80 nm gold nanoparticles. Once the cells and particles were incubated together for 24 h, the labelled products were injected into the rat subretinal layer. Micro-computed tomography was then conducted on the 15th and 30th day post-injection to track the movement of these cells, as visualized by an area of hyperdensity from the coronal section images of the rat head. In addition, we confirmed the cellular uptake of the gold nanoparticles by the mesenchymal stem cells using transmission electron microscopy. As opposed to other methods, the current protocol provides a simple, less labour-intensive and more efficient labelling mechanism for real-time cell tracking. Finally, we discuss the potential manipulations of gold nanoparticles in stem cells for cell replacement and cancer therapy in ocular disorders or diseases.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Mesenchymal Stem Cell Osteogenesis Kit, The Mesenchymal Stem Cell Osteogenesis Kit contains all the reagents necessary to readily differentiate mesenchymal stem cells to an osteogenic lineage as assessed by Alizarin Red staining.
Sigma-Aldrich
Mesenchymal Adipogenesis Kit, Mesenchymal Stem Cell Adipogenesis Kit typically obtains 30% mature adipocytes from the rat bone marrow derived mesenchymal stem cells.