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Three-dimensional cell culture environment promotes partial recovery of the native corneal keratocyte phenotype from a subcultured population.

Abstract
Corneal disease is the fourth leading cause of blindness. According to the World Health Organization, roughly 1.6 million people globally are blind as a result of this disease. The only current treatment for corneal opacity is a corneal tissue transplant. Unfortunately, the demand for tissue exceeds supply, making a tissue-engineered in vitro cornea highly desirable. For an in vitro cornea to be useful, it must be transparent, which requires downregulation of the light-scattering intracellular protein alpha-smooth muscle actin (αSMA) and upregulation of the native corneal marker, aldehyde dehydrogenase 1A1 (ALDH1A1). This study focuses on the effects of a three-dimensional (3D) matrix on the expression levels of αSMA and ALDH1A1 by a subcultured population of rabbit corneal keratocytes and the comparison of the 3D matrix effects to other culture conditions. We show that, through western blot and quantitative real-time PCR, the presence of collagen strongly downregulates αSMA. Further, 3D cultures maintain low actin expression even in the presence of a proinflammatory cytokine, transforming growth factor-beta (TGF-β). Finally, 3D culture conditions show a partial recovery of ALDH1A1 expression, which has never been previously observed in a serum-exposed subcultured cell population. Overall, this study suggests that 3D culture is not only a relatively stronger signal than both collagen and TGF-β, it is also sufficient to induce some recovery of ALDH1A1 and the native corneal phenotype despite the presence of serum.
AuthorsRussell E Thompson, Liana C Boraas, Miranda Sowder, Marta K Bechtel, Elizabeth J Orwin
JournalTissue engineering. Part A (Tissue Eng Part A) Vol. 19 Issue 13-14 Pg. 1564-72 (Jul 2013) ISSN: 1937-335X [Electronic] United States
PMID23410050 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
Chemical References
  • Aldehyde Dehydrogenase
Topics
  • Aldehyde Dehydrogenase (metabolism)
  • Animals
  • Cell Culture Techniques (methods)
  • Cells, Cultured
  • Corneal Keratocytes (cytology, metabolism)
  • Rabbits

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