Manoochehrabadi Tahereh, Samadikuchaksaraei Ali, Solouki Amin, Daryabari Seyed-Hashem, Ghasemi Hamed, Lotfi Ehsan, Mansourian Sajad, Majidi Jila, Milan Peiman Brouki, Gholipourmalekabadi Mazaher
Cellular and Molecular Research Center, Iran University of Medical Sciences, Tehran, Iran.
Department of Tissue Engineering & Regenerative Medicine, Faculty of Advanced Technologies in Medicine, Iran University of Medical Sciences, Tehran, Iran.
Iran J Basic Med Sci. 2025;28(7):916-928. doi: 10.22038/ijbms.2025.86110.18601.
Substrate engineering is one of the attractive fields of changing cell behavior and fate, especially for stem cell (SC) therapies. The SC pool is an essential factor in transplantation outcomes. Here, the objective was to preserve the stemness of the cornea's limbal epithelial stem cell (LESC) using naturally biomimicking corneal cell topography.
A cell-imprinted substrate was prepared using the natural topography of rabbit cornea's LESC. The LESC cells were characterized by immunostaining (ABCG2 and Cytokeratin-12), then re-cultivated on a topography mold (imprinted PDMS), on FLAT PDMS (without any pattern), and the control group (tissue culture plate). 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 .
The 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 and -12 expression (as differentiation-related genes). studies showed a more significant number of cells and the expression of the ABCG2 gene in the imprinted PDMS group. In contrast, higher expressions of the ∆Np63 gene and more stratification were observed in the control group (no treatment). Histological studies showed that the imprinted PDMS group had normal morphology with fully organized collagens.
The results of LESC cultured on imprinted PDMS suggested that LESC cell imprinting could be an excellent substrate for LESC expansion and preserve their stemness for cell therapy.
基质工程是改变细胞行为和命运的一个有吸引力的领域，特别是对于干细胞（SC）治疗。干细胞库是移植结果的一个重要因素。在此，目的是利用天然仿生角膜细胞拓扑结构来保持角膜缘上皮干细胞（LESC）的干性。
利用兔角膜缘上皮干细胞的天然拓扑结构制备细胞印记基质。通过免疫染色（ABCG2和细胞角蛋白-12）对角膜缘上皮干细胞进行表征，然后将其重新培养在拓扑模具（印记聚二甲基硅氧烷）、平整聚二甲基硅氧烷（无任何图案）和对照组（组织培养板）上。最终，在兔角膜上建立碱烧伤模型，并检查细胞印记模具作为角膜伤口愈合植入物的有效性。
结果表明，与其他两组相比，印记聚二甲基硅氧烷使角膜缘上皮干细胞保持干性状态，∆NP63和ABCG2基因（干性相关基因）高表达，细胞角蛋白-3和-12表达低（作为分化相关基因）。研究表明，印记聚二甲基硅氧烷组中的细胞数量和ABCG2基因表达更显著。相比之下，在对照组（未治疗）中观察到∆Np63基因表达更高且分层更多。组织学研究表明，印记聚二甲基硅氧烷组形态正常，胶原组织完整。
在印记聚二甲基硅氧烷上培养角膜缘上皮干细胞的结果表明，角膜缘上皮干细胞印记可能是角膜缘上皮干细胞扩增的优良基质，并能保持其干性用于细胞治疗。
