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Home / Equine Research Bank / Vet Ophthalmol / Effects of growth factors (EGF, PDGF-BB and TGF-beta 1) on c...
Veterinary ophthalmology2003; 6(3); 211-217; doi: 10.1046/j.1463-5224.2003.00296.x

Effects of growth factors (EGF, PDGF-BB and TGF-beta 1) on cultured equine epithelial cells and keratocytes: implications for wound healing.

Abstract: The physiologic mechanisms involving growth factors, including PDGF-BB, EGF, and TGF-beta 1, as potent mediators of fibroblasts and epithelial cells in corneal wound healing remain unknown. The goal of this study was to determine culture methods for equine epithelial cells and keratocytes and to investigate how exogenous growth factors influence proliferation of both cell types. Methods: Cell cultures were established from healthy corneas harvested from horses immediately following euthanasia and maintained using standard tissue culture protocols. To determine the effects of PDGF-BB, EGF, TGF-beta 1, keratocytes (1 x 10(5)/well) and epithelial cells (2 x 10(5)/well) were each cultured in 12 well plates and exposed separately to the growth factors. The cells were exposed to concentrations of EGF between 0 and 50 ng/mL; PDGF-BB between 0 and 75 ng/mL; and TGF-beta 1 between 0 and 10 ng/mL. Cell proliferation was measured using 3H-thymidine assay and differences in growth determined using anova and Tukey's HSD test (P < 0.05). Results: Epithelial cell and keratocyte cultures were successfully established. EGF maximally stimulated keratocyte and epithelial cells at 25 ng/mL and 5 ng/mL, respectively. PDGF-BB maximally stimulated keratocytes and epithelial cells at 50 ng/mL and 5 ng/mL, respectively. TGF-beta 1 inhibited keratocytes at 5 ng/mL and 10 ng/mL, and epithelial cells at 1 ng/mL and 2 ng/mL. Conclusions: Methods were established to maintain epithelial cells and keratocytes in vitro. PDGF-BB and EGF stimulate, while TGF-beta 1 inhibits the proliferation of epithelial cells and keratocytes. These growth factors may play a role in maintenance and repair of the equine cornea.
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Publication Date: 2003-09-03 PubMed ID: 12950652DOI: 10.1046/j.1463-5224.2003.00296.xGoogle Scholar: Lookup
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  • Journal Article
  • Research Support
  • Non-U.S. Gov't
  • Research Support
  • U.S. Gov't
  • P.H.S.

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

This research examines the effects of certain growth factors on horse epithelial cells and keratocytes (cells found in the cornea). The study hopes to improve understanding about how these growth factors can impact wound healing in the cornea.

Establishing Cell Culture Methods

  • The researchers first determined successful methods for culturing equine epithelial cells and keratocytes. This involved harvesting cells from healthy horse corneas post-euthanasia and developing them using standard tissue culture processes.

Growth Factors Exposure and Proliferation Assessment

  • Next, the cells were exposed separately to various concentrations of three growth factors: EGF, PDGF-BB, and TGF-beta 1. The epithelial cells and keratocytes were placed in 12 well plates at a count of 2 x 10(5)/well and 1 x 10(5)/well, respectively.
  • Exposure concentrations ranged as follows: EGF between 0-50 ng/mL; PDGF-BB between 0-75 ng/mL; and TGF-beta 1 between 0-10 ng/mL.
  • To gauge growth factor effects on cells under these conditions, researchers used a 3H-thymidine assay to measure cell proliferation. For determining any differences in cell growth, they utilized statistical methods – anova and Tukey’s HSD test.

Results from Growth Factors Exposure

  • The three growth factors showed differing effects. EGF stimulated maximal cell proliferation at 25 ng/mL for keratocytes and 5 ng/mL for epithelial cells.
  • PDGF-BB showed its maximum stimulation at 50 ng/mL for keratocytes and 5 ng/mL for epithelial cells.
  • Meanwhile, TGF-beta 1 inhibited growth at 5 ng/mL and 10 ng/mL for keratocytes and inhibited epithelial cells at 1 ng/mL and 2 ng/mL.

Conclusions of the Study

  • The establishment of successful methods for culturing equine epithelial cells and keratocytes constitutes an important part of this study. They found that growth factors EGF and PDGF-BB stimulated growth, while TGF-beta 1 inhibited proliferation.
  • The observed effects on cell growth suggest that these growth factors could play a role in corneal repair and maintenance in horses.

Cite This Article

APA
Haber M, Cao Z, Panjwani N, Bedenice D, Li WW, Provost PJ. (2003). Effects of growth factors (EGF, PDGF-BB and TGF-beta 1) on cultured equine epithelial cells and keratocytes: implications for wound healing. Vet Ophthalmol, 6(3), 211-217. https://doi.org/10.1046/j.1463-5224.2003.00296.x

Publication

Veterinary ophthalmology
ISSN: 1463-5216
NlmUniqueID: 100887377
Country: England
Language: English
Volume: 6
Issue: 3
Pages: 211-217

Researcher Affiliations

Haber, Marion
  • Department of Clinical Sciences, Tufts University School of Veterinary Medicine, North Grafton, MA, USA.
Cao, Zhiyi
    Panjwani, Noorjahan
      Bedenice, Daniela
        Li, William W
          Provost, Patricia J

            MeSH Terms

            • Animals
            • Becaplermin
            • Cell Culture Techniques
            • Cell Division / drug effects
            • Cornea / cytology
            • Cornea / drug effects
            • Cornea / physiology
            • Dose-Response Relationship, Drug
            • Epidermal Growth Factor / pharmacology
            • Epithelium, Corneal / drug effects
            • Epithelium, Corneal / physiology
            • Growth Substances / pharmacology
            • Horses / physiology
            • Platelet-Derived Growth Factor / pharmacology
            • Proto-Oncogene Proteins c-sis
            • Transforming Growth Factor beta / pharmacology
            • Transforming Growth Factor beta1
            • Wound Healing

            Grant Funding

            • T35 DK07635 / NIDDK NIH HHS

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