Fabrication of Three-Dimensional Cell Constructs Using Temperature-Responsive Hydrogel

Abstract

A morphologically controlled three-dimensional (3D) cell construct composed of only cells and having no scaffold material might be a valuable biologic material for tissue engineering applications, as the scaffold materials can cause delay of tissue regeneration in some conditions. To obtain such a 3D cell construct, a 3D thermoresponsive hydrogel (poly-N-isopropylacrylamide) was prepared as a mold material that changes its volume depending on the temperature. Three-dimensional osteoblast cell constructs with a variety of morphologies as well as a monolayered cell sheet were obtained by decreasing the surrounding temperature of the hydrogel designed with a predefined shape and formed by curing in a polymer mold manufactured via 3D printing. The cell sheet or 3D cell constructs detachment resulted from a simple change in the gel volume, not by the surface chemistry of the gel, because the surface hydrophilicity of the gel was maintained over a wide temperature range. These 2D/3D cell constructs have numbers of exciting applications such as cell carriers for tissue regeneration or as model tissues for the biological study.

Comparative Analysis of Mouse-Induced Pluripotent Stem Cells and Mesenchymal Stem Cells During Osteogenic Differentiation In Vitro

“…The low expression of these fibril-forming type 1 collagens by osteogenic iPSCs explains the lower amount of fibril structures in their extracellular spaces relative to osteogenic MSCs as observed in our SEM/ TEM studies. In the present study, osteogenic MSCs highly expressed cartilage-associated collagens [72], such as col 2a1, col 6a2, col 11a1, and col 12a1, which is in accord with previous reports, suggesting intrinsic expression of chondrogenic genes during osteogenesis of bone marrow-derived MSCs [41,73,74]. In contrast, osteogenic iPSCs highly expressed col 4a1 and col 4a2, which encode major constituents of the basement membrane, and col 14a1, which encodes an FACIT (fibrillar-associated collagen with interrupted triple-helix) that is widely expressed in several tissues, including bone [72].…”

Evaluation of methods for pore generation and their influence on physio-chemical properties of a protein based hydrogel

“…Because those materials can serve as an excellent growth substrate for cells many approaches have been made to produce artificial networks for 3D cell growth in order to trigger and alter cellular behavior within those structures [2], [3], [4]. This is not only important because it represents a more realistic environment for cells than standard 2D approaches, but it also offers an enormous amount of new possibilities for the growth of cells, especially for stem cells and tumor models [5].…”

Design of Tetra-arm PEG-crosslinked Thermoresponsive Hydrogel for 3D Cell Culture

“…This is because our PEG-RGDS-PNIPAAm gel does not shrink easily due to bulkiness and hydrophilicity of PEG. 16 Roughly 15 and 7% volume changes could be observed in temperature changes from 25 to 37 C and from 33 to 37 C, respectively. Moreover, the kinetics of the volume change of the gel from 25 to 37 C and from 37 to 25 C (Fig.…”