Speaker
Description
Introduction: Tissue decellularization methods allow obtaining new sources of biological scaffolds obtained from natural tissues and organs1,2. Although decellularized scaffolds can be useful, it is well known that their regenerative potential is significantly improved after recellularization. In the present work, we evaluated the potential of several cell sources to recellularize a xenograft scaffold obtained by porcine sclerocorneal limbus decellularization.
Methodology: Porcine corneal limbi were decellularized with 0.1% SDS and thoroughly washed in PBS. Then, decellularized xenografts were recellularized using three different cell sources: cultured corneal epithelial cells (CEC), human adipose derived stem cells (hADSC) and human umbilical cord Wharton’s Jelly stem cells (hWJSC). To favor recellularization, xenografts were casted in agarose molds. Cell recellularization efficacy, limbal stem cell maintenance markers and extracellular matrix (ECM) components were evaluated by histochemical and immunohistochemical methods after 7, 14 and 21 days of ex vivo development.
Results: We found that the recellularization efficiency was different for each cell source. First, we found that CEC were able to attach to the decellularized xenografts and formed a partially stratified epithelium able to express several cytokeratins and limbal stem cell markers from day 7. When hADSC were used, we found that cells attached to the xenograft from day 14-21 of development, with expression of some of the limbal markers. However, we found that hWJSC were not able to efficiently attach and grow on the decellularized tissues during the 21 days of the study.
Conclusions: These results suggest that the decellularized xenografts used in this study can be efficiently recellularized with CEC and, to a lesser extent, with hADSC. Expression of several key markers of limbal stem cells suggest that recellularized cells are able to adapt to the microenvironment of the decellularized tissue kept in culture. These findings suggest that sclerocorneal limbi generated by decellularization-recellularization could have potential clinical usefulness for the treatment of patients with limbal damage or stem cell deficiency.
83767230387
