In vitro cultivation of porcine limbal stem cells

Abstract

Similarly as in other organ structures stem cells are present in cornea residing basal epithelial layer termed palisade of Voight. A growing interest in allografts and xenografts implies a thorough study of regenerative potentials of these cells, as well as a clear description of their patterns in in vitro tissue cultures to be grafted. Recently we have developed a simple method for cultivation of porcine corneal epithelial stem cells obtained by biopsy from the limbal region. Eight enucleated eyes were obtained from four slaughtered pigs. 5 mm2 samples of limbal epithelium were taken by keratotomy method. The primary cultures of these cells showed phenotypic and morphometric characteristics of porcine corneal epithelial cells following May-Grunwald-Giemsa staining. After 5d of sowing they reached 80% of confluence. With the «Night & Day» lenses a total confluence was achieved 5d earlier in comparison to the cells that were grown in the secondary cultures. Accordingly, the use of porcine limbal stem cells has potentials in veterinary medicine (as novel approach in reparative/regenerative medicine of pets, horses and selected breeds), but also are in accordance with the extensive studies on the potential use of xenografts, mainly swine tissues/organs, in humans.

Keywords

• Stem cells, Porcine limbus, Cultivation.