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Efficient characterisation of human cell–bioceramic interactions in vitro and in vivo by using enhanced GFP-labelled mesenchymal stem cells

Paper ID Volume ID Publish Year Pages File Format Full-Text
12542 799 2005 11 PDF Available
Title
Efficient characterisation of human cell–bioceramic interactions in vitro and in vivo by using enhanced GFP-labelled mesenchymal stem cells
Abstract

Human mesenchymal stem cells (hMSCs) were transfected using four retroviral pseudotypes, amphotropic murine leukemia viruses 4070 (MuLV-10A1), a modification of amphotropic pseudotype 4073 (A71G, Q74K, V139M), gibbon ape leukemia virus (GaLV), or feline endogenous virus (RD114) encoding the neomycin resistance (Neor) gene and enhanced green fluorescent protein (eGFP) as genetic markers. It was observed that the MuLV4073 was the most efficient pseudotype for hMSC transfection. The proliferation and differentiation characteristics of eGFP-labelled hMSCs were not significantly different from control hMSCs. G418 selected eGFP-labelled cells were cultured for 3 weeks on two porous, commercially available calcium phosphate bioceramics, a “synthetic hydroxyapatite” and a “deproteinised bone”, before implantation into NOD/SCID mice for up to 4 weeks. The eGFP-labelled hMSCs could be readily visualised by their intense green fluorescence both in vitro and in vivo. In “synthetic hydroxyapatite” implants the cells remained in a monolayer, whereas in “deproteinised bone” implants mineralised tissues were detected by histology, scanning electron microscopy and energy dispersive X-ray spectrometry. From the results, it is concluded that the use of eGFP-labelled hMSCs is an effective tool to trace the fate of hMSCs and evaluate the interactions between cells and ceramics both in vitro and in vivo. This is of great value in prospective assessments of these cell populations for use in tissue engineering applications.

Keywords
Human mesenchymal stem cells (hMSCs); Retroviral transfection; Enhanced green fluorescent protein (eGFP); Murine leukemia viruses (MuLV); Gibbon ape leukemia virus (GaLV); Feline endogenous virus (RD114); Hydroxyapatite ceramics
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Efficient characterisation of human cell–bioceramic interactions in vitro and in vivo by using enhanced GFP-labelled mesenchymal stem cells
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Publisher
Database: Elsevier - ScienceDirect
Journal: Biomaterials - Volume 26, Issue 29, October 2005, Pages 5790–5800
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us