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

Sung Hwan Moon, Jongil Ju, Soon Jung Park, Daekyeong Bae, Hyung Min Chung, Sang Hoon Lee

Research output: Contribution to journalArticle

33 Citations (Scopus)

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 two-fold 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.

Original languageEnglish
Pages (from-to)5987-5997
Number of pages11
JournalBiomaterials
Volume35
Issue number23
DOIs
Publication statusPublished - 2014 Jul

Keywords

  • Concave microwells
  • Embryoid body
  • Human embryonic stem cell
  • Optimize differentiation
  • Size screening

ASJC Scopus subject areas

  • Bioengineering
  • Ceramics and Composites
  • Biophysics
  • Biomaterials
  • Mechanics of Materials

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