Abstract
Purpose:
In patients with limbal stem cell deficiency, in addition to the loss of the limbal epithelial stem cells, the limbal niche is invariably damaged/destroyed. Therefore, strategies are needed to reconstruct the limbal niche. Mesenchymal type cells have been shown to be an important part of the niche. In this study, we examined whether mesenchymal stem cells (MSC) from the bone marrow (BM) can function as niche support cells.
Methods:
Human bone marrow MSCs were cultured in serum containing media. Primary human corneal epithelial cells were grown in KSFM before being passaged onto plates with BM-MSC. Colony forming efficiency of epithelial cells co-cultured with BM-MSC was examined. Conditioned media from BM-MSC and limbal fibroblasts were compared in terms of their ability to promote epithelial growth. BM-MSC were also grown in decellularized human corneas to examine their growth and differentiation in 3D.
Results:
As reported by others, BM-MSC supported colony formation of corneal epithelial cells, which appeared to be dependent on the passage number of the BM-MSC. Pre-treatment of BM-MSC with mitomycin C reduced their ability to support corneal epithelial cell growth as evident by the effect of conditioned media (non-MMC vs MMC treated) on corneal epithelial cells. BM-MSC proliferated in 3D culture conditions in decellularized human cornea adopting a keratocyte-like phenotype.
Conclusions:
These results indicate that BM-MSC can support the clonal growth of corneal epithelial cells. More studies are needed to determine the optimal growth conditions for their use as limbal niche cells.
Keywords: 721 stem cells •
482 cornea: epithelium •
480 cornea: basic science