A thermoresponsive, microtextured substrate for cell sheet engineering with defined structural organization

Abstract

The proper function of many tissues depends critically on the structural organization of the cells and matrix of which they are comprised. Therefore, in order to engineer functional tissue equivalents that closely mimic the unique properties of native tissues it is necessary to develop strategies for reproducing the complex, highly organized structure of these tissues. To this end, we sought to develop a simple method for generating cell sheets that have defined ECM/cell organization using microtextured, thermoresponsive polystyrene substrates to guide cell organization and tissue growth. The patterns consisted of large arrays of alternating grooves and ridges (50 μm wide, 5 μm deep). Vascular smooth muscle cells cultured on these substrates produced intact sheets consisting of cells that exhibited strong alignment in the direction of the micropattern. These sheets could be readily transferred from patterned substrates to non-patterned substrates without the loss of tissue organization. Ultimately, such sheets will be layered to form larger tissues with defined ECM/cell organization that spans multiple length scales.

Development of thermoresponsive poly(propylene-g-N-isopropylacrylamide) non-woven 3D scaffold for smart cell culture using oxyfluorination-assisted graft polymerisation

“…Where layered or patterned co-culture cell sheets are used, the process requires multiple steps and does not address the need for a structural matrix to enable cell growth in three dimensions. In recent years, some 3D PNIPAAm scaffolds have been developed based on hydrogels, grafted porous 3D membranes and micro-textured PNIPAAm surfaces [7,8]. Additionally a PP-g-PNIPAAm non-woven membrane with adsorbed antibodies has recently been reported in the literature for selective cell separation and enrichment of target cells [9].…”

“…The latter lacks structural and organisational cues for cells since the 2D environment does not replicate the complexity of physiological tissue [7]. Where layered or patterned co-culture cell sheets are used, the process requires multiple steps and does not address the need for a structural matrix to enable cell growth in three dimensions.…”

Dynamic Manipulation of Hydrogels to Control Cell Behavior: A Review

“…Intact tissue-like cell sheets with a preserved ECM 95 can be used individually or can be layered/rolled to create tissues of larger sizes or with a defined laminar organization, an approach known as ''cell sheet engineering.'' 96 These cell sheets could also be patterned and assembled to mimic the microarchitecture of native tissues, crucial for functional tissue regeneration.…”

Concise Review: Mechanotransduction via p190RhoGAP Regulates a Switch Between Cardiomyogenic and Endothelial Lineages in Adult Cardiac Progenitors

“…Kshitiz, Afzal, Kim et al 2001 www.StemCells.com V C AlphaMed Press 2014 topographical cues temporarily to cells [38]. The cue provided by the nanotopographic input matching the cardiac ECM ultrastructure caused an increase in p190RhoGAP abundance leading to an increased formation of cardiomyocyte progenitor cells (Fig.…”