Abstract: Objective To investigate the best method to culture and label vocalfold fibroblasts（VFFs）in vitro. Methods VFFs were cultured by enzyme digestion to observe their growth and morphology under an inverted microscope and labeled with Ad-EGFP.The cells（100 million/0.1ml）were injected into the vocal cords of 12 New Zealand rabbits to observe the growth of VFFs. Results VFFs could be cultured in vitro and showed characteristic morphologyof fibroblasts.Immunochemistry showed positive vimentin expression Ad-EGFP could transfect VFFs.No survived cells were observed on the tissue sections 15 days after injection of VFFs. Conclusion VFFs can be cultured by enzyme digestion.Ad-EGFP is an ideal tracing marker.