Purpose: To investigate the most appropriate media condition for the proliferation and functional maintenance of human corneal endothelial cells (HCECs).
Methods: We cultured HCECs in traditional media (medium A or D) and in stem cell media (medium E or N). The morphology of the cells was evaluated by inverted microscopy. Collagen, type VIII, alpha 2 and sodium-potassium adenosine triphosphatase (Na(+)-K(+) ATPase) expression were analyzed as differentiation markers. Octamer-binding transcription factor 3/4, glial fibrillary acidic protein, nestin and β-catenin expression were evaluated as stem cell associated proteins. The cell proliferation rate was evaluated with a cell counting kit-8 assay. Wound healing assays were also performed. The transendothelial electrical potential difference (TEPD) value was used to estimate the endothelial cell permeability in vitro.
Results: The proliferation and morphology analyses demonstrated that there were significant differences between the media. The expression of differentiation markers and stem cell-associated proteins was different between the media. Medium D resulted in higher proliferation rates compared with the other media, while still maintaining the differentiation potential and surface marker expression profile characteristic of HCECs. Compared with other media, TEPD was higher in medium N.
Conclusions: Culture medium D was superior to the other media with regard to the expression of stem cell-associated proteins, proliferation, and cell migration. However, medium N was more appropriate than the other three media with regard to maintaining the proper cell shape and function.
Keywords: COL8A2; GFAP; Na+-K+ ATPase; OCT3/4; human corneal endothelial cells; media; nestin.
Copyright 2014 The Association for Research in Vision and Ophthalmology, Inc.