Journal
JOURNAL OF TISSUE ENGINEERING AND REGENERATIVE MEDICINE
Volume 2, Issue 8, Pages 499-506Publisher
WILEY
DOI: 10.1002/term.125
Keywords
embryonic stem cells; embryoid body; chondrogenesis; differentiation; tissue engineering; cartilage; sorting
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Funding
- Whitaker Foundation
- Arthritis Foundation
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For applications in tissue engineering and regenerative medicine, embryonic stem cells (ESCs) are commonly pre-differentiated in the form of embryoid bodies (EBs). The uncontrolled cell differentiation in EBs results in a highly heterogeneous cell population, an unfavourable condition for therapeutic development. The purpose of this study was to determine an optimal size of EBs for chondrogenic differentiation. EBs were produced in suspension culture with mouse ESCs (ES-D3 GL). The 5-day-old EBs were sorted under a microscope by diameter: small EBs (S-EBs, <100 mu m), medium EBs (M-EBs, 100-150 mu m) and large EBs (L-EBs, > 150 mu m). The three sizes of EBs were cultured separately for 3 weeks in chondrogenic medium. Type 11 collagen and aggrecan gene expression was significantly upregulated in the S-EBs, when compared with the M-EBs and L-EBs (p < 0.05 and p < 0.001, respectively). Proteoglycans produced by the cells derived from S-EBs were >50% of the other two groups. in addition, both Oct4 and Sox2 were expressed more in S-EBs than in M-EBs and L-EBs. Type X collagen expression was relatively increased in L-EBs. Slight shifts toward haematopoietic and endothelial differentiation were seen in the L- and M-EBs. In summary, the size of EBs has implications on ESC differentiation. Cells derived from S-EBs have a greater chondrogenic potential than those from M-EBs and L-EBs. The size of EBs can be a parameter utilized to optimize ESC differentiation for tissue engineering. Copyright (C) 2008 John Wiley & Sons, Ltd.
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