title: Difference in suitable mechanical properties of three-dimensional, synthetic scaffolds for self-renewing mouse embryonic stem cells of different genetic backgrounds.
abstract: We evaluated whether the genetic background of embryonic stem cells (ESCs) affects the properties suitable for three-dimensional (3D) synthetic scaffolds for cell self-renewal. Inbred R1 and hybrid B6D2F1 mouse ESC lines were cultured for 7 days in hydrogel scaffolds with different properties derived from conjugating 7.5, 10, 12.5, or 15% (wt/vol) vinylsulfone-functionalized three-, four-, or eight-arm polyethylene glycol (PEG) with dicysteine-containing crosslinkers with an intervening matrix metalloproteinase-specific cleavage sites. Cell proliferation and expression of self-renewal-related genes and proteins by ESCs cultured in feeder-free or containing 2D culture plate or 3D hydrogel were monitored. As a preliminary experiment, the E14 ESC-customized synthetic 3D microenvironment did not maintain self-renewal of either the R1 or B6D2F1 ESCs. The best R1 cell proliferation (10.04 vs. 0.16-4.39; p < 0.0001) was observed in the four-arm 7.5% PEG-based hydrogels than those with other properties, whereas the F1 ESCs showed better proliferation when they were embedded in the three-arm 10% hydrogels. Self-renewal-related gene and protein expression by ESCs after feeder-free 3D culture was generally maintained compared with the feeder-containing 2D culture, but expression patterns and quantities differed. However, the feeder-free 3D culture yielded better expression than the feeder-free 2D culture. In conclusion, genetic background determined the suitability of hydrogel scaffolds for self-renewal of ESCs, which requires customization for the mechanical properties of each cell line. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 105B: 2261-2268, 2017.
authors.affiliation: Department of Agricultural Biotechnology, Seoul National University, Seoul, 151-921, Korea.
authors.forename: Myungook
authors.initials: M
authors.lastname: Lee
authors.affiliation: Research Institutes of Agriculture and Life Sciences, Seoul National University, Seoul, 151-921, Korea.
authors.forename: Jong Il
authors.initials: JI
authors.lastname: Ahn
authors.affiliation: Department of Agricultural Biotechnology, Seoul National University, Seoul, 151-921, Korea.
authors.forename: Ji Yeon
authors.initials: JY
authors.lastname: Ahn
authors.affiliation: Department of Agricultural Biotechnology, Seoul National University, Seoul, 151-921, Korea.
authors.forename: Woo Sub
authors.initials: WS
authors.lastname: Yang
authors.affiliation: Institute of Bioengineering (IBI) and Institute of Chemical Sciences and Engineering (ISIC), School of Life Sciences, École Polytechnique Fédérale de Lausanne (EPFL), CH-1015 Lausanne, Switzerland; Institute for Molecular Engineering, University of Chicago, IL 60637, USA.
authors.forename: Jeffrey A
authors.initials: JA
authors.lastname: Hubbell
authors.affiliation: Department of Agricultural Biotechnology, Seoul National University, Seoul, 151-921, Korea.|Research Institutes of Agriculture and Life Sciences, Seoul National University, Seoul, 151-921, Korea.
authors.forename: Jeong Mook
authors.initials: JM
authors.lastname: Lim
authors.affiliation: Department of Animal Life Science and Division of Applied Animal Science, Kangwon National University, Chuncheon, 200-701, Korea.
authors.forename: Seung Tae
authors.initials: ST
authors.lastname: Lee
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D002460:Cell Line; D049109:Cell Proliferation; D005030:Ethylenes / Q000737:chemistry; D005786:Gene Expression Regulation; D020100:Hydrogels / Q000737:chemistry*; D051379:Mice; D000066450:Mouse Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism*; D011092:Polyethylene Glycols / Q000737:chemistry; D013451:Sulfonic Acids / Q000737:chemistry; D054457:Tissue Scaffolds / Q000737:chemistry*
chemical_list: D005030:Ethylenes; D020100:Hydrogels; D013451:Sulfonic Acids; D011092:Polyethylene Glycols; C085726:ethylenesulfonic acid
keywords: embryonic stem cells; genetic background; polyethylene glycol-based hydrogel; self-renewal; three-dimensional synthetic hydrogel scaffold