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Ultrastructural analysis of mouse embryonic stem cell-derived chondrocytes

Abstract

Pluripotent embryonic stem (ES) cells cultivated as cellular aggregates, so called embryoid bodies (EBs), differentiate spontaneously into different cell types of all three germ layers in vitro resembling processes of cellular differentiation during embryonic development. Regarding chondrogenic differentiation, murine ES cells differentiate into progenitor cells, which form pre-cartilaginous condensations in the EB-outgrowths and express marker molecules characteristic for mesenchymal cell types such as Sox5 and Sox6. Later, mature chondrocytes appear which express collagen type II, and the collagen fibers show a typical morphology as demonstrated by electron-microscopical analysis. These mature chondrogenic cells are organized in cartilage nodules and produce large amounts of extracellular proteoglycans as revealed by staining with cupromeronic blue. Finally, cells organized in nodules express collagen type X, indicating the hypertrophic stage. In conclusion, differentiation of murine ES cells into chondrocytes proceeds from the undifferentiated stem cell via progenitor cells up to mature chondrogenic cells, which then undergo hypertrophy. Furthermore, because the ES-cell-derived chondrocytes did not express elastin, a marker for elastic cartilage tissue, we suggest the cartilage nodules to resemble hyaline cartilage tissue.

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Acknowledgements

The skilful technical assistance of A. Eirich and M. Dose is gratefully acknowledged. The work was supported by the Medical Faculty of the University of Lübeck and funded by Intermed Service GmbH&CoKG (Geesthacht, Germany) and Eppendorf AG (Hamburg, Germany).

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Correspondence to Jan Kramer.

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Kramer, J., Klinger, M., Kruse, C. et al. Ultrastructural analysis of mouse embryonic stem cell-derived chondrocytes. Anat Embryol 210, 175–185 (2005). https://doi.org/10.1007/s00429-005-0020-x

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  • DOI: https://doi.org/10.1007/s00429-005-0020-x

Keywords

  • ES cells
  • Chondrocytes
  • Collagen
  • Cartilage
  • Chondrogenic differentiation