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SHR Neuro Cancer Cardio Lipid Metab Microb

Kramer, J; Schlenke, P; Rohwedel, J.
Induction of ES cell-derived cartilage formation.
Curr Protoc Cell Biol. 2007; Chapter 23(6):Unit 23.5-Unit 23.5 Doi: 10.1002/0471143030.cb2305s34
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Co-authors Med Uni Graz: Schlenke Peter
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Abstract:: This unit describes the protocols used for cultivation of murine embryonic stem (ES) cells and their differentiation into chondrogenic cell types in vitro. ES cells cultivated as cellular aggregates, so-called embryoid bodies (EBs), differentiate spontaneously into chondrogenic cell types recapitulating cellular events of chondro- and osteogenesis. The undifferentiated ES cells differentiate into mesenchymal prechondrogenic cells in the EB outgrowths. These progenitor cells aggregate and form mesenchymal condensations. During further cultivation, these cells form cartilage nodules, show a phenotype typical for chondroblasts, and start to express marker molecules of cartilage tissue. Later, the chondrocytes become hypertrophic, and finally, marker molecules indicating bone formation can be detected in the nodules. This unit also contains protocols for characterization of the differentiated cells by immunostaining, mRNA-in situ hybridization, electron microscopy, and RT-PCR analysis.
Find related publications in this database (using NLM MeSH Indexing): Animals -; Biological Markers - analysis; Cell Culture Techniques - instrumentation Cell Culture Techniques - methods; Cell Differentiation -; Cells, Cultured - cytology Cells, Cultured - drug effects; Chondrocytes - cytology; Coloring Agents -; Culture Media - pharmacology; Embryonic Stem Cells - cytology Embryonic Stem Cells - drug effects; Fibroblasts - cytology; Gene Expression Profiling - methods; Immunohistochemistry - methods; In Situ Hybridization, Fluorescence - methods; Mesenchymal Stromal Cells - cytology Mesenchymal Stromal Cells - drug effects; Mice -; Microscopy, Electron - methods; Osteocytes - cytology; Pluripotent Stem Cells - cytology Pluripotent Stem Cells - drug effects; Reverse Transcriptase Polymerase Chain Reaction - methods; Staining and Labeling - methods