A method for identification of vocal fold lamina propria fibroblasts in culture
Received 8 November 2007; received in revised form 25 August 2008; accepted 9 September 2008.
Objective
Vocal fold biology research is emerging as a vital area of study in laryngology. One impediment is the lack of both commercially available vocal fold lamina propria fibroblasts and a constitutively expressed specific marker for fibroblasts. We present an in vitro technique that allows for identification of fibroblasts by ruling out the possibility of the cells belonging to other lineages that are found in vocal fold tissue.
Study Design
An in vitro study.
Methods
Two primary vocal fold fibroblast cell lines and one immortalized vocal fold fibroblast cell line were cultured. Immunohistologic staining for α-actinin, cytokeratin 19, and von Willebrand factor was completed for the three fibroblast lines in addition to skeletal, endothelial, and epithelial cell lines. Cell type was differentiated by positive staining for α-actinin, cytokeratin 19, and von Willebrand factor.
Results
Fibroblast cultures did not express α-actinin, cytokeratin 19, and von Willebrand factor, whereas skeletal muscle, endothelial, and epithelial cultured cells expressed each respectively.
Conclusions
This simple rule-out methodology for fibroblast confirmation is an important step when establishing cell culture, and it establishes sound internal validity particularly in the early stages of this emerging area of study.
Department of Surgery, Division of Otolaryngology–Head and Neck Surgery, University of Wisconsin Madison, Madison, WI; and Department of Surgery, Division of Otolaryngology–Head and Neck Surgery, The University of Utah, Salt Lake City, UT
Corresponding author: Susan L. Thibeault, PhD, Department of Surgery, Division of Otolaryngology–Head and Neck Surgery, University of Wisconsin Madison, Madison, WI
ABSTRACT