title: Effects of Culture Dimensions on Maintenance of Porcine Inner Cell Mass-Derived Cell Self-Renewal.
abstract: Despite the fact that porcine embryonic stem cells (ESCs) are a practical study tool,in vitrolong-term maintenance of these cells is difficult in a two-dimensional (2D) microenvironment using cellular niche or extracellular matrix proteins. However, a three-dimensional (3D) microenvironment, similar to that enclosing the inner cell mass of the blastocyst, may improvein vitromaintenance of self-renewal. Accordingly, as a first step toward constructing a 3D microenvironment optimized to maintain porcine ESC self-renewal, we investigated different culture dimensions for porcine ICM-derived cells to enhance the maintenance of self-renewal. Porcine ICM-derived cells were cultured in agarose-based 3D hydrogel with self-renewal-friendly mechanics and in 2D culture plates with or without feeder cells. Subsequently, the effects of the 3D microenvironment on maintenance of self-renewal were identified by analyzing colony formation and morphology, alkaline phosphatase (AP) activity, and transcriptional and translational regulation of self-renewal-related genes. The 3D microenvironment using a 1.5% (w/v) agarose-based 3D hydrogel resulted in significantly more colonies with stereoscopic morphology, significantly improved AP activity, and increased protein expression of self-renewal-related genes compared to those in the 2D microenvironment. These results demonstrate that self-renewal of porcine ICM-derived cells can be maintained more effectively in a 3D microenvironment than in a 2D microenvironment. These results will help develop novel culture systems for ICM-derived cells derived from diverse species, which will contribute to stimulating basic and applicable studies related to ESCs.
authors.affiliation: Department of Animal Life Science, Kangwon National University, Chuncheon 24341, Korea.
authors.forename: Song
authors.initials: S
authors.lastname: Baek
authors.affiliation: Department of Animal Life Science, Kangwon National University, Chuncheon 24341, Korea.
authors.forename: Na Rae
authors.initials: NR
authors.lastname: Han
authors.affiliation: Division of Animal Resource Science, Kangwon National University, Chuncheon 24341, Korea.
authors.forename: Jung Im
authors.initials: JI
authors.lastname: Yun
authors.affiliation: Department of Cellular and Molecular Physiology, Yale School of Medicine, New Haven 06510, USA.
authors.forename: Jae Yeon
authors.initials: JY
authors.lastname: Hwang
authors.affiliation: Animal Nutrition and Physiology Team, National Institute of Animal Science, RDA, Wanju 55365, Korea.
authors.forename: Minseok
authors.initials: M
authors.lastname: Kim
authors.affiliation: Department of Animal Life Science, Kangwon National University, Chuncheon 24341, Korea.|Division of Applied Animal Science, Kangwon National University, Chuncheon 24341, Korea.
authors.forename: Choon Keun
authors.initials: CK
authors.lastname: Park
authors.affiliation: College of Veterinary Medicine, Kangwon National University, Chuncheon 24341, Korea.
authors.forename: Eunsong
authors.initials: E
authors.lastname: Lee
authors.affiliation: Department of Animal Life Science, Kangwon National University, Chuncheon 24341, Korea.|Division of Applied Animal Science, Kangwon National University, Chuncheon 24341, Korea.
authors.forename: Seung Tae
authors.initials: ST
authors.lastname: Lee
mesh_terms: D000818:Animals; D053624:Blastocyst Inner Cell Mass / Q000166:cytology*; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation / Q000502:physiology; D049109:Cell Proliferation / Q000502:physiology; D053595:Embryonic Stem Cells / Q000166:cytology*; D005260:Female; D008813:Mice, Inbred ICR; D011247:Pregnancy; D013552:Swine
keywords: agarose; culture dimension; embryonic stem cells; pig; self-renewal