January 1992
Volume 33, Issue 1
Free
Articles  |   January 1992
Clonal growth and differentiation of rabbit meibomian gland epithelium in serum-free culture: differential modulation by EGF and FGF.
Author Affiliations
  • S L Maskin
    Department of Ophthalmology, Bascom Palmer Eye Institute, University of Miami School of Medicine, Florida.
  • S C Tseng
    Department of Ophthalmology, Bascom Palmer Eye Institute, University of Miami School of Medicine, Florida.
Investigative Ophthalmology & Visual Science January 1992, Vol.33, 205-217. doi:
  • Views
  • PDF
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      S L Maskin, S C Tseng; Clonal growth and differentiation of rabbit meibomian gland epithelium in serum-free culture: differential modulation by EGF and FGF.. Invest. Ophthalmol. Vis. Sci. 1992;33(1):205-217.

      Download citation file:


      © ARVO (1962-2015); The Authors (2016-present)

      ×
  • Supplements
This content is PDF only. Please click on the PDF icon to access.
Abstract

We have established a serum-free clonal culture system to study the growth and differentiation of individual progenitor epithelial cells of the meibomian gland independent of other cell types and undefined serum factors. Single meibomian gland epithelial cells were obtained by subjecting whole meibomian glands to a brief EDTA treatment, needle aspiration, and nylon mesh filtration. The cells had been isolated by a previously described method using enzymatic and microsurgical techniques. Four to five hundred cells obtained were seeded on a 35 mm or 60 mm dish with serum-free MCDB 151 medium containing insulin, transferrin, selenium, ethanolamine, o-phosphorylethanolamine, dimethyl sulfoxide, and calcium. Control cultures with this basic medium did not show continuous clonal growth. Addition of epidermal growth factor (EGF) from 1 to 5 and 10 ng/ml enhanced clonal growth with a decreasing effect as measured by colony forming efficacy and colony size. Clonal growth was associated with the occurrence of two types of colony morphology and an increase in intracellular lipid production in the BrdU-labelled cells, shown by Nile red fluorescent staining. In contrast, the clonal growth stimulated by addition of acidic fibroblast growth factor (aFGF) from 1 to 100 ng/ml exhibited a pattern of an initial increase followed by a decrease, with the maximum noted at 10 ng/ml. Moreover, the aFGF-stimulated clonal growth was associated with a uniform colony morphology and minimal lipid synthesis even in the non-BrdU-labelled cells. These results indicate that clonal growth of meibomian gland epithelium can be achieved in a serum-free culture by adding either of these two peptide growth factors. Furthermore, the clonal growth stimulated by EGF was associated with progressive cellular differentiation more so than that of aFGF. Further exploration of such a differential regulation of proliferation and differentiation by EGF and aFGF may provide a better understanding of normal meibomian gland function and pathogenesis of various meibomian gland disorders.

×
×

This PDF is available to Subscribers Only

Sign in or purchase a subscription to access this content. ×

You must be signed into an individual account to use this feature.

×