Effects of Bone Marrow Mesenchymal Stem Cells on Cell Proliferation and Growth Factor Expression of Limbal Epithelial Cells in vitro

Abstract

Mesenchymal stem cells (MSCs) can play an active role in the reconstruction of the ocular surface; however, the mechanisms by which the implanted MSCs exert their effects remain elusive. In the present study, we investigated the influences of MSCs on cell proliferation and growth factor expression of corneal limbal epithelial cells (LECs) in vitro. Rat LECs were either cocultured with rat MSCs or cultured in rat MSC-conditioned medium. The harvested LECs were subjected to light microscopy, MTT assay, bromodeoxyuridine (BrdU) staining, flow cytometry and real-time quantitative RT-PCR analysis, respectively. Light microscopy showed that coculturing of LECs with MSCs significantly increased the proliferation of LECs. The MTT assay showed that MSC-conditioned medium significantly increased the cell viability of LECs compared to those incubated with plain medium. BrdU immunostaining and flow-cytometric analysis of the cell cycle showed that the cell proliferation rate of LECs cocultured with MSCs was significantly higher than that of LECs cultured alone. Real-time quantitative RT-PCR analysis showed that expression of epidermal growth factor in LECs cocultured with MSCs was significantly higher than that in LECs cultured alone. However, expression of basic fibroblast growth factor showed no difference under the two culture conditions. These results indicate that MSCs promote the survival and proliferation of LECs, and these effects may be mediated in a paracrine manner.

Therapeutic Effect of Human Adipose Tissue-Derived Mesenchymal Stem Cells in Experimental Corneal Failure Due to Limbal Stem Cell Niche Damage

“…Our results, together with those obtained by other authors, indicate that MSCs contribute to the recovery of the corneal epithelium by secreting factors that act at a paracrine level rather than by a transdifferentiation process. These factors would reduce neovascularization and inflammation and provide an enabling environment to promote the proliferation and differentiation of the resident stem cells in the tissues . Although the anti‐angiogenic and the anti‐inflammatory capacity of hAT‐MSCs have been reported , this is the first time that the capability of these cells to recover the corneal and limbal phenotypes has been demonstrated in in vivo models.…”

Mesenchymal stem cells: Potential role in corneal wound repair and transplantation

“…The survival and proliferation of rat LECs are markedly promoted and the expression of EGF is upregulated by co-culturing LECs and rat MSCs or by culturing LECs in a medium pre-conditioned with rat MSCs. The effects of MSCs on LECs might be mediated by paracrine action [64] . In summary, paracrine mechanisms of MSCs may exert a significant impact in promoting corneal wound repair, which involves the joint participation of different soluble factors that modulate inflammation and angiogenesis as well as improve tissue regeneration.…”

Progenitor Cells for Ocular Surface Regenerative Therapy

“…When epithelial sheets produced by both cocultures were transplanted onto chemically damaged rabbit corneas, a similarly robust expression of CK3 was observed without corneal neovascularization or conjunctivalization [50]. More recently, the ability of BM-MSCs to promote LSC proliferation and influence improvements in cell viability and growth factor expression was reported in cocultures and in the presence of LSC-conditioned media [51]. These data also confirmed the suitability of BM-MSCs for supporting ex vivo expansion of LSCs.…”