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Optimizing human embryonic stem cells differentiation efficiency by screening size-tunable homogenous embryoid bodies

Authors
Moon, Sung-HwanJu, JongilPark, Soon-JungBae, DaekyeongChung, Hyung-MinLee, Sang-Hoon
Issue Date
7월-2014
Publisher
ELSEVIER SCI LTD
Keywords
Human embryonic stem cell; Concave microwells; Size screening; Embryoid body; Optimize differentiation
Citation
BIOMATERIALS, v.35, no.23, pp.5987 - 5997
Indexed
SCIE
SCOPUS
Journal Title
BIOMATERIALS
Volume
35
Number
23
Start Page
5987
End Page
5997
URI
https://scholar.korea.ac.kr/handle/2021.sw.korea/98114
DOI
10.1016/j.biomaterials.2014.04.001
ISSN
0142-9612
Abstract
Human embryonic stem cells (hESCs) are generally induced to differentiate by forming spherical structures termed embryoid bodies (EBs) in the presence of soluble growth factors. hEBs are generated by suspending small clumps of hESC colonies; however, the resulting hEBs are heterogeneous because this method lacks the ability to control the number of cells in individual EBs. This heterogeneity affects factors that influence differentiation such as cell cell contact and the diffusion of soluble factors, and consequently, the differentiation capacity of each EB varies. Here, we fabricated size-tunable concave microwells to control the physical environment, thereby regulating the size of EBs formed from single hESCs. Defined numbers of single hESCs were forced to aggregate and generate uniformly sized EBs with high fidelity, and the size of the EBs was controlled using concave microwells of different diameters. Differentiation patterns in H9- and CHA15-hESCs were affected by EB size in both the absence and presence of growth factors. By screening EB size in the presence of various BMP4 concentrations, a twofold increase in endothelial cell differentiation was achieved. Because each hESC line has unique characteristics, the findings of this study demonstrate that concave microwells could be used to screen different EB sizes and growth factor concentrations to optimize differentiation for each hESC line. (C) 2014 Elsevier Ltd. All rights reserved.
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