Comparison of ex vivo cultivated human limbal epithelial stem cell viability and proliferation on different substrates

Abstract

Ocular surface injury causes serious vision-related problems especially when limbal stem cells are affected. Treatment lies in the transplantation of viable donor cells. Various substrates are used for the cultivation of limbal epithelial stem cells. In the present study, viability and proliferation of ex vivo cultured limbal epithelial stem cells were examined on a variety of substrates like collagen type IV, direct plastic Petri plate, intact amniotic membrane and denuded amniotic membrane. Viability and proliferation of cells were examined by colorimetric assay and [(3)H]-thymidine incorporation study. Furthermore, matrix metalloproteinase is known to be a key regulator in stem cell migration and proliferation. This enzyme activity was studied by gelatinolytic zymography. It was found from this study that although human limbal epithelial stem cells could be cultivated on different substrates such as collagen type IV, direct plastic Petri plate, intact amniotic membrane and denuded amniotic membrane, maximum growth and proliferation was observed when cultured on intact amniotic membrane. The number of patients suffering from limbal epithelial stem cell deficiency is large compared to donor tissues available for transplantation. Hence, increased cell viability and proliferation is required to serve more patients.