Abstract
Induced pluripotent stem cells (iPSC) are a promising cell source for cartilage regenerative medicine; however, the methods for chondrocyte induction from iPSC are currently developing and not yet sufficient for clinical application. Here, we report the establishment of a fluorescent indicator system for monitoring chondrogenic differentiation from iPSC to simplify screening for effective factors that induce chondrocytes from iPSC. We generated iPSC from embryonic fibroblasts of Col2a1-EGFP transgenic mice by retroviral transduction of Oct4, Sox2, Klf4, and c-Myc. Among the 30 clones of Col2a1-EGFP iPSC we established, two clones showed high expression levels of embryonic stem cell (ESC) marker genes, similar to control ESC. A teratoma formation assay showed that the two clones were pluripotent and differentiated into cell types from all three germ layers. The fluorescent signal was observed during chondrogenic differentiation of the two clones concomitant with the increase in chondrocyte marker expression. In conclusion, Col2a1-EGFP iPSC are useful for monitoring chondrogenic differentiation and will contribute to research in cartilage regenerative medicine.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Cell Differentiation / genetics
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Cell Differentiation / physiology
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Cells, Cultured
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Collagen Type II / genetics
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Collagen Type II / metabolism
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Embryonic Stem Cells / cytology
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Embryonic Stem Cells / metabolism
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Green Fluorescent Proteins / genetics
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Green Fluorescent Proteins / metabolism
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Induced Pluripotent Stem Cells / cytology*
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Induced Pluripotent Stem Cells / metabolism*
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Kruppel-Like Factor 4
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Kruppel-Like Transcription Factors / genetics
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Kruppel-Like Transcription Factors / metabolism
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Mice
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Mice, Transgenic
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Octamer Transcription Factor-3 / genetics
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Octamer Transcription Factor-3 / metabolism
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Proto-Oncogene Proteins c-myb / genetics
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Proto-Oncogene Proteins c-myb / metabolism
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SOXB1 Transcription Factors / genetics
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SOXB1 Transcription Factors / metabolism
Substances
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Col2a1 protein, mouse
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Collagen Type II
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Klf4 protein, mouse
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Kruppel-Like Factor 4
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Kruppel-Like Transcription Factors
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Octamer Transcription Factor-3
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Proto-Oncogene Proteins c-myb
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SOXB1 Transcription Factors
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Sox2 protein, mouse
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enhanced green fluorescent protein
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Green Fluorescent Proteins
Grants and funding
This work was supported by Grants-in-Aid for Scientific Research and the Project for Realization of Regenerative Medicine from the Ministry of Education, Science, Sports, Culture and Technology (MEXT) of Japan. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.