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Originally published In Press as doi:10.1167/iovs.08-2857 on August 6, 2009
 (Investigative Ophthalmology and Visual Science. 2010;51:192-200.)
© 2010 by The Association for Research in Vision and Ophthalmology, Inc.
doi:10.1167/iovs.08-2857
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Perimysial Fibroblasts of Extraocular Muscle, as Unique as the Muscle Fibers

Linda L. Kusner,1,2,3 Andrew Young,4 Steven Tjoe,4 Patrick Leahy,5 and Henry J. Kaminski3

From the Departments of 1Ophthalmology, 2Pharmacological and Physiological Science, and 3Neurology and Psychiatry, Saint Louis University, St. Louis, Missouri; and the 4Department of Neurology, 5Case Comprehensive Cancer Center, Case Western Reserve University, Cleveland, Ohio.

Corresponding author: Linda L. Kusner, Department of Ophthalmology, Saint Louis School of Medicine, St. Louis, MO 63104; lkusner{at}slu.edu.

Purpose. Extraocular muscle (EOM) has a distinct skeletal muscle phenotype. The hypothesis for the study was that fibroblasts support the unique EOM phenotype and that perimysial fibroblasts derived from EOM have properties that distinguish them from fibroblasts derived from other skeletal muscle.

Methods. Perimysial fibroblasts from leg muscle (LM-Fibro) and EOM (EOM-Fibro) of mice were derived and maintained in culture. EOM- and LM-Fibro were assessed morphologically and for vimentin, smooth muscle actin, and Thy-1 immunoreactivity. DNA microarray analysis was performed on LM- and EOM-Fibro grown in conditions that support myoblast differentiation. To assess trophic interactions, co-cultures of myoblasts from established cell lines, CL-EOM and CL-LM with, EOM- or LM-Fibro were performed in direct contact and in a permeable filter support culture. The degree of myotube maturation was assessed by the percentage of myotubes with more than three myonuclei per myotube.

Results. EOM- and LM-Fibro cells exhibited distinct morphologies. Both cell types proliferated as a monolayer and expressed vimentin. Fifty-five percent (SD 4.4%) of EOM-Fibro were Thy-1 positive compared with only 24% (SD 4.4%) of LM-Fibro. DNA microarray analysis demonstrated differential expression of structural, immune response, and metabolism-related genes between EOM- and LM-Fibro. Co-cultures demonstrated that mature myotube formation in EOM-derived cell lines was supported to a greater extent by EOM-Fibro than by LM-Fibro, compared with CL-EOM grown with LM-Fibro.

Conclusions. Fibroblasts from EOM demonstrate distinct properties that distinguish them from leg muscle–derived fibroblasts. The distinct properties of EOM-Fibro may support the unique EOM phenotype and contribute to their differential involvement in disease.






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