Anatomical location and culture of equine corneal epithelial stem cells

Hidekazu Moriyama,Atsutoshi Kuwano,Yoshinori Kasashima,Shinya Wada

doi:10.1111/vop.12050

Hidekazu Moriyama, Atsutoshi Kuwano + Show 2 more

Open Access

https://doi.org/10.1111/vop.12050

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Abstract

To identify morphologically the locations of equine corneal epithelial stem cells (CESCs) and to culture these cells. We studied the eyes of 12 adult thoroughbred horses. Eye tissues were immunostained for two positive stem cell markers (p63, CK14) and one negative marker (CK3) to identify the locations of CESCs, so we could compare their immunostaining patterns with those of human stem cells previously reported. We compared the proliferation rates and morphological features of epithelial cells isolated from the corneal limbus and central cornea. Undifferentiated cells expressing the same immunostaining pattern as human CESCs were present in the equine corneal limbus. Cultured epithelial cells isolated from the limbus expressed the same immunostaining pattern that CESCs show histologically, but cells isolated from the central cornea did not proliferate and could not be evaluated. Equine CESCs were localized in the epithelial basal layer of the corneal limbus, where melanocytes reside. They could be cultured without loss of their undifferentiated nature. When collecting such stem cells, it may be useful to harvest and culture corneal epithelial tissues in the limbus where melanocytes serve as an indicator of the collecting area.

Highlights

Racehorses have a high incidence of corneal injuries because their proximity to each other when racing can result in trauma
Localization of corneal epithelial stem cells (CESCs) in the equine corneal limbus Eyeballs were collected from adult thoroughbred horses that were being killed in a slaughterhouse and were immediately transported to our laboratory in phosphatebuffered saline
Localization of CESCs in the equine corneal limbus In sections stained for p63, the nuclei of 2–3 layers of corneal epithelial basal cells located apically from the basolateral side stained positive, consistent with the region where melanocytes are present in the corneal limbus, (Fig. 2b,B)

Summary

Results

Undifferentiated cells expressing the same immunostaining pattern as human CESCs were present in the equine corneal limbus. Cultured epithelial cells isolated from the limbus expressed the same immunostaining pattern that CESCs show histologically, but cells isolated from the central cornea did not proliferate and could not be evaluated

Conclusions

INTRODUCTION

MATERIALS AND METHODS

RESULTS

DISCUSSION