Dynamic culture of mesenchymal stem cells on thermo-responsive PNIPAM substrate

Abstract

Event Abstract Back to Event Dynamic culture of mesenchymal stem cells on thermo-responsive PNIPAM substrate George Altankov1 and Maria Bianchi2 1 ICREA and Institute for Bioengineering of Catalonia, Molecular Dynamics at Cell-Biomaterials Interface, Spain 2 Institute for Bioengineering of Catalonia, Molecular Dynamics at Cell-Biomaterials Interface, Spain As of today, stem cell-based therapies are still hindered by the insufficient knowledge of the signals that govern stem cell behavior. Stem cell niche is a dynamic microenvironment that balances stem cells activity to maintain tissue homeostasis and repair. The development of strategies to mimic this dynamic niche environment provides insight to our understanding how to control the behavior of stem cells. In this work we used smart thermo-responsive polymer poly(N-isopropylacrylamide (PNIPAM) for culturing of adipose derived mesenchymal stem cells (ADSCs) applying periodic temperature cycles to perturb the cell-substratum interactions. We further studied the functional behavior of stem cells following their two principal properties: the capability for self-renewal (i.ee. the symmetric growth) and to differentiate (asymmetric cell division). Material and Methods: 2.0×104 cells/well ADSCs of 2nd passage were cultured on regular tissue culture polystyrene (control) or on thermo-responsive polymer (PNIPAAm) substrate (Nunc UpCell™) and further submitted to equal thermo-cycling consisting of 20 min cooling (37ºC to 24ºC) and 20 min heating (24ºC to 37ºC), twice per day using specially designed thermo-plate with circulating water jacked fitted to a standard CO2 incubator. For the assessment of symmetric cell division the cells were propagated in basal medium with passaging at each 3rd day and counted before seeding again for the next passage in the initial density. The cumulative population doubling level (CPDL) was calculated and compared for up to 13th passage monitoring also the overall cell morphology and fibronectin matrix secretion as indirect measure for ADSCs ageing. For assessing the asymmetric cell growth the ADSC were switched to osteogenic differentiation medium and again subjected to dynamic thermo-cycling condition. The kinetics of cells growth, alkaline phosphatase activity (AP) and Ca deposition (Alisarin red) were assayed at 3, 7, 14 and 21 dais. Results and Discussion: This is the first study identifying in vitro the effect of thermos-cycling (2 cycles per day) on the behaviour of ADSCs on PNIPAM substrate. The general conclusion was that periodic alterations in the adhesive phenotype of ADSCs significantly alter their differentiation potential but not the symmetric cells renewal in an undifferentiated state. This work was supported by CIBER-BBN (Spain) and the European Commission through the FP7 Industry-Academia Partnerships and Pathways (IAPP) project FIBROGELNET; The valuable support of the project MAT2012-38359-C03-03 HEALINSYNERGY, funded by Spanish Ministry of Science and Innovation is also acknowledged. Keywords: Cell Adhesion, Cell Differentiation, stem cell, Smart material Conference: 10th World Biomaterials Congress, Montréal, Canada, 17 May - 22 May, 2016. Presentation Type: Poster Topic: Adhesive biomaterials Citation: Altankov G and Bianchi M (2016). Dynamic culture of mesenchymal stem cells on thermo-responsive PNIPAM substrate. Front. Bioeng. Biotechnol. Conference Abstract: 10th World Biomaterials Congress. doi: 10.3389/conf.FBIOE.2016.01.02120 Copyright: The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers. They are made available through the Frontiers publishing platform as a service to conference organizers and presenters. The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated. Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed. For Frontiers’ terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions. Received: 27 Mar 2016; Published Online: 30 Mar 2016. Login Required This action requires you to be registered with Frontiers and logged in. To register or login click here. Abstract Info Abstract The Authors in Frontiers George Altankov Maria Bianchi Google George Altankov Maria Bianchi Google Scholar George Altankov Maria Bianchi PubMed George Altankov Maria Bianchi Related Article in Frontiers Google Scholar PubMed Abstract Close Back to top Javascript is disabled. Please enable Javascript in your browser settings in order to see all the content on this page.