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Mol Cells. 2017 Feb;40(2):117-122. doi: 10.14348/molcells.2017.2223. Epub 2017 Feb 15.

Effects of Culture Dimensions on Maintenance of Porcine Inner Cell Mass-Derived Cell Self-Renewal.

Author information

1
Department of Animal Life Science, Kangwon National University, Chuncheon 24341, Korea.
2
Division of Animal Resource Science, Kangwon National University, Chuncheon 24341, Korea.
3
Department of Cellular and Molecular Physiology, Yale School of Medicine, New Haven 06510, USA.
4
Animal Nutrition and Physiology Team, National Institute of Animal Science, RDA, Wanju 55365, Korea.
5
Division of Applied Animal Science, Kangwon National University, Chuncheon 24341, Korea.
6
College of Veterinary Medicine, Kangwon National University, Chuncheon 24341, Korea.

Abstract

Despite the fact that porcine embryonic stem cells (ESCs) are a practical study tool, in vitro long-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 improve in vitro maintenance 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.

KEYWORDS:

agarose; culture dimension; embryonic stem cells; pig; self-renewal

PMID:
28196411
PMCID:
PMC5339502
DOI:
10.14348/molcells.2017.2223
[Indexed for MEDLINE]
Free PMC Article
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