Skeletal muscle fiber-specific green autofluorescence: potential for stem cell engraftment artifacts.
Adult stem cell research has lately been plagued by controversy regarding the possibility that some adult stem cells can engraft into nonautochthonous tissues. While most reports have observed some level of engraftment, the prevalence has varied in some cases by two orders of magnitude, suggesting that major technical variations may underlie these differences, possibly outweighing the biological basis of the observations. Here we describe bright green autofluorescence in a specific subset of skeletal muscle fibers that strongly resembles emission from green fluorescent protein (GFP). Moreover, we show that oxidative muscle fibers exhibit this autofluorescence, likely due to flavin, associated with NADH dehydrogenase. Finally, we demonstrate that confocal microscopy, in conjunction with spectral scanning, can be used to distinguish between GFP and autofluorescence. We suggest this autofluorescence artifact may account for some of the discrepancies in this field, particularly those describing skeletal muscle engraftment.
Center for Cell and Gene Therapy, Department of Pediatrics, Baylor College of Medicine, Houston, Texas, USA.
Kathyjo A
KA
Jackson
D Scott
DS
Snyder
Margaret A
MA
Goodell
D000818:Animals; D016477:Artifacts; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000378:metabolism*; D051379:Mice; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D018485:Muscle Fibers, Skeletal / Q000378:metabolism / Q000637:transplantation*; D018482:Muscle, Skeletal / Q000293:injuries / Q000378:metabolism / Q000637:transplantation*; D009218:Myosins / Q000378:metabolism; D009245:NADH Dehydrogenase / Q000378:metabolism*; D033581:Stem Cell Transplantation / Q000379:methods
D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins; D009245:NADH Dehydrogenase; D009218:Myosins