In vitro differentiation of mouse embryonic stem cells: Enrichment of endodermal cells in the embryoid body

Dongho Choi, Hye Ja Lee, Seunghyun Jee, Soojung Jin, Soo Kyung Koo, Seung Sam Paik, Sung Chul Jung, Sue Yun Hwang, Kwang Soo Lee, Bermseok Oh

Research output: Contribution to journalArticlepeer-review

59 Scopus citations

Abstract

Embryonic stem (ES) cells have the potential to differentiate into all three germ layers, providing new perspectives not only for embryonic development but also for the application in cell replacement therapies. Even though the formation of an embryoid body (EB) in a suspension culture has been the most popular method to differentiate ES cells into a wide range of cells, not much is known about the characteristics of EB cells. To this end, we investigated the process of EB formation in the suspension culture of ES cells at weekly intervals for up to 6 weeks. We observed that the central apoptotic area is most active in the first week of EB formation and that the cell adhesion molecules, except β-catenin, are highly expressed throughout the examination period. The sequential expression of endodermal genes in EBs during the 6-week culture correlated closely with that of normal embryo development. The outer surface of EBs stained positive for α-fetoprotein and GATA-4. When isolated from the 2-week-old EB by trypsin treatment, these endodermal lineage cells matured in vitro into hepatocytes upon stimulation with various hepatotrophic factors. In conclusion, our results demonstrate that endodermal cells can be retrieved from EBs and matured into specific cell types, opening new therapeutic usage of these in vitro differentiated cells in the cell replacement therapy of various diseases.

Original languageEnglish
Pages (from-to)817-827
Number of pages11
JournalStem Cells
Volume23
Issue number6
DOIs
StatePublished - Jun 2005

Keywords

  • Differentiation
  • Embryoid body
  • Embryonic stem cell
  • Endoderm

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