Culture and characterization of human dental pulp‑derived stem cells as limbal stem cells for corneal damage repair

Abstract

limbal stem cell deficiency (lScd) is one of the leading causes of corneal damage. Injury or inflammation in the cornea causes lScd, which may be unilateral or bilateral depending upon the cause. limbal epithelial cell implants successfully improve vision in patients with chemical injury-induced lScd. Transplantation of cultured epithelial stem cells has become a treatment of choice for numerous patients with lScd. Bilateral lScd is frequently observed in the general population, where no residual stem cells are available for ex vivo culture. allografts are associated with a high risk of rejection, neoplasia, and disease transmission. in this respect, allogenic cell populations from other regions in the patient may substitute for allogenic material. in the present study, dental pulp stem cells were cultured in limbal stem cell media and these cells were characterized against limbal stem cells, revealing the significance of using dental pulp stem cell treatment in bilateral lScd. The morphology and culture pattern of both limbal and dental pulp stem cells grown in limbal stem-specific media were similar. Polymerase chain reaction analysis revealed that stem cell markers were highly expressed in limbal stem cells compared to in dental pulp stem cells, regardless of the medium and scaffold in which they were grown. although dental pulp stem cell molecular expression is quite low at the transcript level, the functional protein level according to immunocytochemistry and western blot analyses demonstrated that stem cells and corneal differentiation molecule levels were quite high, indicating their potential as limbal stem cells in the respective microenvironment.

Profile of biological characterizations and clinical application of corneal stem/progenitor cells

“…Alternative cell populations: Compared with traditional transplants, the application of alternative cell populations avoids corneal donor shortages and the risk of disease transmission, graft rejection and tumours (due to immunosuppressant application) associated with allografts, which makes it possible to replace traditional transplants. Currently, the alternative cell populations include oral mucosal epithelial cells[ 135 , 136 ], human embryonic stem cells (HESCs)[ 137 ], induced pluripotent stem cells (iPSCs)[ 138 , 139 ], mesenchymal stem cells (MSCs)[ 140 , 141 ], human immature dental pulp stem cells[ 142 , 143 ], etc. Among these alternatives, MSCs and iPSCs are of vital importance and will be described in detail.…”

Goals and Challenges of Stem Cell-Based Therapy for Corneal Blindness Due to Limbal Deficiency

“…The transplantation of these stem cells into a LSCD rabbit model reduced corneal neovascularization and conjunctivalization, and reconstructed the damaged ocular surface by developing a well-formed corneal epithelium that expressed limbal epithelial stem cell and corneal epithelial cell markers [ 197 , 198 ]. These and other authors further developed the capacity of dental pulp stem cells to acquire corneal and limbal epithelial features using different cell culture techniques [ 199 , 200 ] and cell carriers [ 201 , 202 ]. However, although these cells seem to represent a valid alternative source of cells for treating patients with LSCD, more preclinical evidence should be gathered, especially related to their tumorigenicity, before translating this technology into clinical practice.…”