objective: to study the effects of different matrices and co-culture systems on cultured limbal stem cells (lscs). materials and methods: limbal explants were co-cultured with limbal fibroblasts (lf) and/or mouse embryonic fibroblasts (mef) on filter inserts coated with amniotic membrane (am), matrigel (mat) and collagen type i (col). results: this study revealed that am facilitated the cell mi...

Human limbal epithelial cells (HLE) and corneal stromal stem cells (CSSC) reside in close proximity in vivo in the corneal limbal stem cell niche. However, HLE are typically cultured in vitro without supporting niche cells. Here, we re-create the cell-cell juxtaposition of the native environment in vitro, to provide a tool for investigation of epithelial-stromal cell interactions and to optimiz...

PURPOSE To determine the relative expression of cell cycle-association proteins in human corneal and limbal epithelium and corneal endothelium in situ, to correlate staining patterns of cell cycle-associated proteins with known proliferative status of corneal and limbal epithelial cells, and to determine the relative proliferative status of corneal endothelial cells in situ by comparing their s...

We investigated the corneal morphology of adult Mp/+ mice, which are heterozygous for the micropinna microphthalmia mutation, and identified several abnormalities, which implied that corneal epithelial maintenance was abnormal. The Mp/+ corneal epithelium was thin, loosely packed and contained goblet cells in older mice. Evidence also suggested that the barrier function was compromised. However...

PURPOSE To examine the expression of stage-specific embryonic antigen-4 (SSEA4) in the epithelium of the human ocular surface and characterize SSEA4(+) and SSEA4(-) limbal epithelial cells. METHODS SSEA4 expression in the human cornea and limbus was examined by RT-PCR and immunohistochemistry. SSEA4(+) and SSEA4(-) cells were then separated by using magnetic beads. The phenotypes of these two...

PURPOSE Simple limbal epithelial transplantation (SLET) and cultivated limbal epithelial transplantation (CLET) are proven clinical techniques for treating limbal stem cell deficiency (LSCD). However, the ideal size and number of the limbal explants required for transplantation has not been clearly elucidated. This in vitro study aimed to determine the optimal limbal explant size required for c...

PURPOSE To investigate the expression and role of the Wnt signaling pathway in human limbal stem cells (LSCs). METHODS Total RNA was isolated from the human limbus and central cornea. Limbal or cornea-specific transcripts were identified through quantitative real-time PCR. Protein expression of Wnt molecules was confirmed by immunohistochemistry on human ocular tissue. Activation of Wnt signa...

AIMS The aim of this work was to investigate different surgical options for the repair of the ocular surface, using various extensions of the procedure of limbal stem cell allotransplantation. METHODS/RESULTS Straightforward lamellar limbal transplantation was performed in one patient with contact lens induced limbal stem cell failure. A second patient with a neoplastic corneal lesion underwe...

Corneal epithelial stem cells are thought to be localized in the basal cell layer of the limbus. We developed a monoclonal antibody designated 4G10.3 that immunolocalized to limbal basal cells in rat corneas. Western blot analysis demonstrated that 4G10.3 reacted with a single band of 50,000 molecular weight in rat and rabbit corneal epithelium and 48,000-49,000 in human epithelium. Following e...