Effective generation of autologous corneal keratocytes from mesenchymal stem cells by cell revTM mSC diffKera for clinical use

Abstract

Background & Aim Autologous corneal tissue regeneration is an ideal source of cornea transplants as it minimizes the risks of inflammation and immune rejection. Corneal blindness is one of the two leading causes of blindness worldwide. Currently, cornea transplantation is at the mercy of donated corneas which limited numbers in many countries result in more than 5 years waiting-list for such operative therapy. Because mesenchymal Stem Cells(mSC) are armed with key features of multipotent differentiation potentials, and they are also a feasible source to obtain from patients, here by using our Cell RevTM msc diffKera recipe, we effectively generate autologous corneal keratocytes, the critical corneal stromal cells, needed to maintain corneal structure and function. Methods, Results & Conclusion Our formulation includes FDA approved biochemical components essential to direct and maintain corneal keratocyte cell fate. Within 7 days of culturing mSCs in our diffKera media, we were able to achieve 5 folds proliferation as well as efficient differentiation such that we observed more than 90% dendritic star-like morphology similar to keratocytes but not mSCs. Like keratocytes, the newly differentiated cells were positive to proteoglycans production specific to corneal stroma, lumican, as well as keratocyte biomarkers such as ALDH1A1 and CD34. The control media, dubbed Keratocyte media (KM), commonly used in maintaining keratocytes as well as differentiating mSCs to keratocytes, showed only 20% cell growth and only a few dendritic shaped cells. Our generated keratocytes also expressed key components to corneal function i.e., collagen transcripts, and importantly, they did not express alpha smooth actin nor high levels of TGF-beta as these proteins are parts of inflammation signaling. These results suggest that our mSC diffKera formulation provides potential solution to effective autologous corneal keratocytes, and thus bringing us one step closer to autologous cornea generation in the near future.