Efficient mesenchymal stem cell conversion to corneal keratocytes by the use of specific biomaterial and Cell Rev™ mSC diffKera media

Background & Aim Corneal blindness is a prevalent cause of vision loss, affecting approximately 4.9 million people worldwide (Flaxman et al, 2017). The demand for donor replacement corneas often exceeds the supply, requiring alternative treatment options. To achieve corneal function while minimizing the risks of donor rejection, we propose an autologous biomimetic cornea whereby patient-derived mesenchymal stem cells (MSCs) are converted to corneal-like cells on scaffolds that closely resemble the structure of the human cornea. We hypothesize that the effectiveness of cell differentiation depends on the biomaterials used and the cultured media composition. The more closely the environment resembles that of the human cornea, the higher the likelihood that a greater proportion of MSCs will differentiate into corneal keratocytes. Methods, Results & Conclusion Here, we investigated MSC differentiation into corneal keratocytes using various medium formulations on our electrospun scaffolds. Polycaprolactone-bovine dermis type I atelocollagen (PCL-atelocollagen) and polycaprolactone (PCL) solutions were electrospun to obtain the aligned nanofibrous scaffolds. MSCs were cultured on the scaffolds for 12 days in 3 different types of media: the expansion media described by Lynch & Ahearne (2007); keratocyte differentiation media used in the same study; and our Cell Rev™ mSC diffKera media. Immunofluorescence staining for keratocyte markers such as aldehyde dehydrogenase 1 and lumican was conducted on cells fixed on the scaffolds at days 6 and 12. The results confirmed that a greater proportion of MSCs differentiated into keratocytes in Cell Rev™ mSC diffKera media compared with the media used by Lynch & Ahearne (Figure 1). In addition, electrospun PCL-atelocollagen scaffolds are likely to support keratocyte differentiation better than electrospun PCL scaffolds (control) because corneal stromal collagen fibrils comprise mainly type I collagen. These findings demonstrate that the use of our electrospun PCL-atelocollagen scaffolds and Cell Rev™ mSC diffKera formulation effectively enhances MSC differentiation into keratocytes, and represents a novel step for the creation of artificial corneas.