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Toward Corneal Limbus In Vitro Model: Regulation of hPSC-LSC Phenotype by Matrix Stiffness and Topography During Cell Differentiation Process
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A1 Alkuperäisartikkeli tieteellisessä aikakauslehdessä
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en
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16
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Advanced Healthcare Materials, Volume 12, issue 29
Abstract
A functional limbal epithelial stem cells (LSC) niche is a vital element in the regular renewal of the corneal epithelium by LSCs and maintenance of good vision. However, little is known about its unique structure and mechanical properties on LSC regulation, creating a significant gap in development of LSC-based therapies. Herein, the effect of mechanical and architectural elements of the niche on human pluripotent derived LSCs (hPSC-LSC) phenotype and growth is investigated in vitro. Specifically, three formulations of polyacrylamide gels with different controlled stiffnesses are used for culture and characterization of hPSC-LSCs from different stages of differentiation. In addition, limbal mimicking topography in polydimethylsiloxane is utilized for culturing hPSC-LSCs at early time point of differentiation. For comparison, the expression of selected key proteins of the corneal cells is analyzed in their native environment through whole mount staining of human donor corneas. The results suggest that mechanical response and substrate preference of the cells is highly dependent on their developmental stage. In addition, data indicate that cells may carry possible mechanical memory from previous culture matrix, both highlighting the importance of mechanical design of a functional in vitro limbus model.
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Funding Information: The authors would like to acknowledge the Tampere CellTech Laboratories, the Biocenter Finland (BF) and Tampere Imaging Facility (TIF) for their service. They also thanked D. Sc. Laura Koivusalo and Dr. Jan Bruder for assistance in whole mount imaging and laboratory technicians Outi Melin and Hanna Pekkanen for hPSC‐LSC production. The study was funded by Academy of Finland, Graduate school of Faculty of Medicine and Health technology, Tampere University, Finnish Cultural Foundation, Instrumentarium Foundation, the Eye and Tissue Bank Foundation. Publisher Copyright: © 2023 The Authors. Advanced Healthcare Materials published by Wiley-VCH GmbH.
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Kauppila, M, Mörö, A, Valle-Delgado, J J, Ihalainen, T, Sukki, L, Puistola, P, Kallio, P, Ilmarinen, T, Österberg, M & Skottman, H 2023, 'Toward Corneal Limbus In Vitro Model: Regulation of hPSC-LSC Phenotype by Matrix Stiffness and Topography During Cell Differentiation Process', Advanced Healthcare Materials, vol. 12, no. 29, 2301396. https://doi.org/10.1002/adhm.202301396