Differentiation of mesenchymal stem cells into keratocytes by cell imprinting method
Mahsa Fallah Tafti 1 , Hossein Aghamollaei2 , Shahab Faghihi1 , Mehrdad Moosazadeh Moghadam3 , Khosrow Jadidi4 *
- Stem Cell and Regenerative Medicine Group, National Institute of Genetic Engineering and Biotechnology, Tehran 14965/161, Iran
- Chemical Injuries Research Center, Systems Biology and Poisonings Institute, Baqiyatallah University of Medical Sciences, Tehran, Iran
- Applied Biotechnology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran
- Chemical Injures Research Center. Systems Biology and Poisonings Institute, Baqiyatallah University of Medical Sciences, Tehran, Iran.
Abstract: Visual impairments and blindness can accompany corneal stromal disorders such as keratoconus, trauma, and ocular burns. They can be associated with a loss of corneal stromal keratocytes (CSKs). Reconstructing stromal tissue with CSK replacement can be a viable alternative to corneal transplantation, which is restricted by the global donor material shortage and graft failure. Human CSKs are ordinarily quiescent; they proliferate but lose their characteristic phenotype and become stromal fibroblasts. Therefore, due to the sources of human keratocytes being limited for clinical application, the differentiation of stem cells into keratocyte cells is considered for corneal stromal regeneration. It has been demonstrated that mimicking micromechanical forces or micro/nanotopographical environments to control the stem cells' fate can be effective. Here, the potency of substrates with imprinted cell-like topographies for direct and physical differentiation of adipose-derived mesenchymal stem cells (ADSCs) into keratocytes is reported.
Methods: The substrates with imprinted keratocyte-like topographies are prepared. Keratocytes are isolated from the discarded human corneas of Descemet's stripping endothelial keratoplasty (DSEK) surgery. The obtained cells are grown in an enriched media culture and fixed. Then their morphologies are transferred to polydimethylsiloxane (PDMS) substrates by mold casting. Subsequently, mesenchymal stem cells (MSCs) are seeded on the CSK-imprinted substrates. Their differentiation into keratocytes is evaluated by immunocytochemistry at duration times of 14 and 21 days.
Results: Analysis of morphology and expression of CSK-specific markers shows that MSCs cultured on the imprinted substrates have the typical CSK-like morphology and express specific markers of keratocytes, including keratocan and lumican.
Conclusion: It is believed that cell-imprinting is an applicable method to induce the specific cell-like topographies and differentiation of ADSCs into keratocyte cells directly. This physical differentiation method can be proposed .for corneal regeneration and cell therapy.
