RRC ID 68711
Author Saito T, Yano F, Mori D, Ohba S, Hojo H, Otsu M, Eto K, Nakauchi H, Tanaka S, Chung UI, Kawaguchi H.
Title Generation of Col2a1-EGFP iPS cells for monitoring chondrogenic differentiation.
Journal PLoS One
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.
Volume 8(9)
Pages e74137
Published 2013-1-1
DOI 10.1371/journal.pone.0074137
PII PONE-D-13-10240
PMID 24066106
PMC PMC3774617
MeSH Animals Cell Differentiation / genetics Cell Differentiation / physiology Cells, Cultured Collagen Type II / genetics Collagen Type II / metabolism Embryonic Stem Cells / cytology Embryonic Stem Cells / metabolism Green Fluorescent Proteins / genetics Green Fluorescent Proteins / metabolism Induced Pluripotent Stem Cells / cytology* Induced Pluripotent Stem Cells / metabolism* Kruppel-Like Factor 4 Kruppel-Like Transcription Factors / genetics Kruppel-Like Transcription Factors / metabolism Mice Mice, Transgenic Octamer Transcription Factor-3 / genetics Octamer Transcription Factor-3 / metabolism Proto-Oncogene Proteins c-myb / genetics Proto-Oncogene Proteins c-myb / metabolism SOXB1 Transcription Factors / genetics SOXB1 Transcription Factors / metabolism
IF 2.74
Human and Animal Cells mouse embryonic stem cells