Green fluorescent protein is a stable morphological marker for schwann cell transplants in bioengineered nerve conduits.
Bioengineered systems incorporate cultured cells to mimic the substituted tissue. A labeling method is necessary to monitor the survival of transplanted cells within the host. This labeling method must be compatible with the histochemical methods used for morphological analysis. This study assessed (1) The in vitro characteristics of Schwann cells (SCs) labeled with green fluorescent protein (GFP), (2) the in vivo effect of transplanted GFP-SCs in a model of peripheral nerve injury, and (3) the compatibility of GFP-SCs with immunofluorescence histochemical techniques. SCs were retrovirally labeled with GFP and their growth characteristics were compared with those of nontransduced SCs (ntSCs). GFP-SCs were seeded in a resorbable nerve conduit for grafting into a 1-cm gap in rat sciatic nerve. Grafts were harvested after 2 weeks and immunofluorescent staining was performed to measure axonal and SC regeneration distances and to identify GFP-SCs. Results of GFP-SC vitality assays did not vary significantly from those of ntSC assays. GFP-SCs were readily located ex vivo and stimulated significantly better axonal and SC regeneration distances in comparison with empty conduits. These findings show that GFP labeling does not have a deleterious effect on SCs and that it is a useful labeling method for the study of bioengineered systems.
Tohill
Mel P
MP
Blond McIndoe Research Laboratories, University of Manchester, Manchester, UK.
Mann
Dawn J
DJ
Mantovani
Cristina M
CM
Wiberg
Mikael
M
Terenghi
Giorgio
G
D000818:Animals; D000831:Animals, Newborn; D015415:Biomarkers / Q000378:metabolism; D001698:Biomedical Engineering / Q000379:methods; D002470:Cell Survival / Q000502:physiology*; D002478:Cells, Cultured; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism*; D008856:Microscopy, Fluorescence / Q000379:methods; D009416:Nerve Regeneration / Q000502:physiology; D051381:Rats; D017207:Rats, Sprague-Dawley; D011994:Recombinant Proteins / Q000378:metabolism; D012583:Schwann Cells / Q000378:metabolism / Q000473:pathology* / Q000637:transplantation*; D012584:Sciatic Nerve / Q000293:injuries / Q000378:metabolism / Q000473:pathology* / Q000601:surgery*; D023822:Tissue Engineering / Q000379:methods*; D019737:Transplants
D015415:Biomarkers; D011994:Recombinant Proteins; D049452:Green Fluorescent Proteins