title: Fabrication of Patterned Thermoresponsive Microgel Strips on Cell-Adherent Background and Their Application for Cell Sheet Recovery.
abstract: Interfaces between materials and cells play a critical role in cell biomedical applications. Here, a simple, robust, and cost-effective method is developed to fabricate patterned thermoresponsive poly(N-isopropylacrylamide-co-styrene) microgel strips on a polyethyleneimine-precoated, non-thermoresponsive cell-adherent glass coverslip. The aim is to investigate whether cell sheets could be harvested from these cell-adherent surfaces patterned with thermoresponsive strips comprised of the microgels. We hypothesize that if the cell-to-cell interaction is strong enough to retain the whole cell sheet from disintegration, the cell segments growing on the thermoresponsive strips may drag the cell segments growing on the cell-adherent gaps to detach, ending with a whole freestanding and transferable cell sheet. Critical value concerning the width of the thermoresponsive strip and its ratio to the non-thermoresponsive gap may exist for cell sheet recovery from this type of surface pattern. To obtain this critical value, a series of strip patterns with various widths of thermoresponsive strip and non-thermoresponsive gap were prepared using negative microcontact printing technology, with COS7 fibroblast cells being used to test the growth and detachment. The results unraveled that COS7 cells preferentially attached and proliferated on the cell-adherent, non-thermoresponsive gaps to form patterned cell layers and that they subsequently proliferated to cover the microgel strips to form a confluent cell layer. Intact COS7 cell sheets could be recovered when the width of the thermoresponsive strip is no smaller than that of the non-thermoresponsive gap. Other cells such as HeLa, NIH3T3, 293E, and L929 could grow similarly; that is, they showed initial preference to the non-thermoresponsive gaps and then migrated to cover the entire patterned surface. However, it was difficult to detach them as cell sheets due to the weak interactions within the cell layers formed. In contrast, when COS7 and HeLa cells were cultured successively, they formed the cocultured cell layer that could be detached together. These freestanding patterned cell sheets could lead to the development of more elaborate tumor models for drug targeting and interrogation.
authors.affiliation: State Key Laboratory of Heavy Oil Processing and Centre for Bioengineering and Biotechnology, China University of Petroleum (East China) , Qingdao 266580, China.
authors.forename: Yongqing
authors.initials: Y
authors.lastname: Xia
authors.affiliation: State Key Laboratory of Heavy Oil Processing and Centre for Bioengineering and Biotechnology, China University of Petroleum (East China) , Qingdao 266580, China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Tang
authors.affiliation: State Key Laboratory of Heavy Oil Processing and Centre for Bioengineering and Biotechnology, China University of Petroleum (East China) , Qingdao 266580, China.
authors.forename: Han
authors.initials: H
authors.lastname: Wu
authors.affiliation: Biological Physics Laboratory, School of Physics and Astronomy, University of Manchester , Schuster Building, Oxford Road, Manchester M13 9PL, United Kingdom.
authors.forename: Jing
authors.initials: J
authors.lastname: Zhang
authors.affiliation: Biological Physics Laboratory, School of Physics and Astronomy, University of Manchester , Schuster Building, Oxford Road, Manchester M13 9PL, United Kingdom.
authors.forename: Zongyi
authors.initials: Z
authors.lastname: Li
authors.affiliation: Biological Physics Laboratory, School of Physics and Astronomy, University of Manchester , Schuster Building, Oxford Road, Manchester M13 9PL, United Kingdom.
authors.forename: Fang
authors.initials: F
authors.lastname: Pan
authors.affiliation: State Key Laboratory of Heavy Oil Processing and Centre for Bioengineering and Biotechnology, China University of Petroleum (East China) , Qingdao 266580, China.
authors.forename: Shengjie
authors.initials: S
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Heavy Oil Processing and Centre for Bioengineering and Biotechnology, China University of Petroleum (East China) , Qingdao 266580, China.
authors.forename: Xiaojuan
authors.initials: X
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Heavy Oil Processing and Centre for Bioengineering and Biotechnology, China University of Petroleum (East China) , Qingdao 266580, China.
authors.forename: Hai
authors.initials: H
authors.lastname: Xu
authors.affiliation: Biological Physics Laboratory, School of Physics and Astronomy, University of Manchester , Schuster Building, Oxford Road, Manchester M13 9PL, United Kingdom.
authors.forename: Jian Ren
authors.identifier: http://orcid.org/0000-0001-5648-3564
authors.initials: JR
authors.lastname: Lu
mesh_terms: D000818:Animals; D002448:Cell Adhesion; D002460:Cell Line; D006801:Humans; D051379:Mice; D013499:Surface Properties; D013696:Temperature
keywords: biointerface; cell sheet harvesting; functional interface; microgel strips; micropatterning; thermoresponsive cell removal; thermoresponsive microgels