Substrate engineering using naturally biomimicking corneal cell topography for keeping stemness of corneal limbal epithelial-stem cells
Seyed Hashem Daryabari1 , Tahereh Manoochehrabadi2 *, Ali Samadikuchaksaraei3 , Amin Solouki4 , Hamed Ghasemi5 , Ehsan Lotfi3 , Sajad Mansourian5 , Jila Majidi2 , Mazaher Gholipourmalekabadi2
- Baghiatollah University of Medical Sciences, Tehran, Islamic Republic of Iran
- Cellular and Molecular Research Center, Iran University of Medical Sciences, Tehran, Iran,
- Department of Medical Biotechnology, Faculty of Allied Medicine, Iran University of Medical Sciences, Tehran, Iran
- Department of Hematology and Blood Banking, School of Allied Medical Sciences, Shahid Beheshti University of Medical Sciences, Tehran, Iran
- Eye Research Center, Farabi Eye Hospital, Tehran University of Medical Sciences, Tehran, Iran
Abstract: Substrate engineering is one of the attractive fields of changing cell behavior and fate.
Methods: Here, a cell-imprinted substrate was prepared using the natural topography of rabbit cornea's limbal epithelial stem cell (LESC). The LESC cells were re-cultivated on a topography mold (Imprinted PDMS), on FLAT PDMS, and the control group. Ultimately, an alkaline burn model was created on a rabbit's cornea, and the effectiveness of cell-imprinted molds as implants for healing corneal wounds was examined in a live setting.
Results: The in vitro results showed that Imprinted PDMS kept LESC cells in a state of stemness with high expression of ?NP63 and ABCG2 genes (stemness-associated genes) compared to the other two groups and low Cytokeratin 3, 12 expression (as differentiation-related genes). In vivo studies showed a more significant number of cells and the expression of the ABCG2 gene in the Imprinted PDMS group, while higher expressions of the ?Np63 gene and more stratification were observed in the control group (no treatment). Histological studies showed the Imprinted PDMS group had normal morphology with fully organized collagens.
Conclusion: The results suggested that LESC cell imprinting could be an excellent substrate for LESC expansion and preserve their stemness for cell therapy.
