Abstract
Purpose: :
The use of stem cells is promising for future cell-based therapies such as tissue regeneration and engineering. Mesenchymal stem cells (MSCs) derived from adult human adipose tissue (PLA cells) manifest multilineage differentiation capacity. However, their plasticity to differentiate into epithelial characteristics still remains little well-known. The corneal epithelium is maintained by limbal stem cells. Transplantation of autologous corneal stem cells it is not possible in several cases in which bilateral disease produces total corneal stem-cell deficiency. For this reason, the use of autologous MSCs derived from adult human adipose tissue as a source of cells for the ocular surface reconstruction is being studied.
Methods: :
The clonogenic ability of MSCs, as well as assays of adhesion and proliferation within different matrices, has been studied. Cell population distribution has been also compared. Different markers have been analyzed by immunostaining and flow cytometry methods. Human corneal epithelial cells (HCE) were used as control for specific markers.
Results: :
According to adhesive proprieties, two subpopulations (fast and slow adhesion cells) of adult MSCs were isolated from lipoaspirates. Differential patterns of adhesion and proliferation and low colony-formation efficiency were observed. Some differences were shown in adhesion and proliferation assays using several extracellular matrix proteins as coating. In slow adhesion cells (SN2), type I collagen and fibronectin improved adhesion whereas type IV collagen and laminin association improved proliferation state. Laminin presented a negative selection action. CD34, CD90, keratin 3/12 and vimentin were positive for both subpopulations. E-cadherin, keratin 1/5/10/14, p63 and ABCG2 were negative or scantly positive.
Conclusions: :
Adult MSCs could be a potential cell type for ocular surface regenerative medicine, and the expression of keratin 3/12 in PLA cells subpopulations may support the hypothesis of its differentiation to corneal epithelial-like cells.
Keywords: cornea: epithelium • differentiation • regeneration