Tabriz University of Medical Sciences Advanced Pharmaceutical Bulletin 2228-5881 12 2 2022 03 09 The Combined Thermoresponsive Cell-Imprinted Substrate, Induced Differentiation, and "KLC Sheet" Formation 356 365 10.34172/apb.2022.034 EN Neda Keyhanvar https://orcid.org/0000-0001-6852-9865 Nosratollah Zarghami Alexander Seifalian Peyman Keyhanvar Rana Sarvari Roya Salehi Reza Rahbarghazi Mohammadreza Ranjkesh Molood Akbarzadeh Mahdi Mahdipour Mohammad Nouri https://orcid.org/0000-0002-5367-9956 Journal Article 10.34172/apb.2022.034 2020 07 12 2021 05 01 Purpose: Stem cells can exhibit restorative effects with the commitment to functional cells.Cell-imprinted topographies provide adaptable templates and certain dimensions for thedifferentiation and bioactivity of stem cells. Cell sheet technology using the thermo-responsivepolymers detaches the "cell sheets" easier with less destructive effects on the extracellularmatrix (ECM). Here, we aim to dictate keratinocyte-like differentiation of mesenchymal stemcells (MSCs) by using combined cell imprinting and sheet technology.Methods: We developed the poly dimethyl siloxane (PDMS) substrate having keratinocytecell-imprinted topography grafted with the PNIPAAm polymer. Adipose tissue-derived MSCs(AT-MSCs) were cultured on PDMS substrate for 14 days and keratinocyte-like differentiationmonitored via the expression of involucrin, P63, and cytokeratin 14.Results: Data showed the efficiency of the current protocol in the fabrication of PDMSmolds. The culture of AT-MSCs induced typical keratinocyte morphology and up-regulatedthe expression of cytokeratin-14, Involucrin, and P63 compared to AT-MSCs cultured on theplastic surface (P<0.05). Besides, KLC sheets were generated once slight changes occur in theenvironment temperature.Conclusion: These data showed the hypothesis that keratinocyte cell imprinted substrate canorient AT-MSCs toward KLCs by providing a specific niche and topography.