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Original Article |
1 Wisconsin National Primate Research Center, University of Wisconsin, Madison, WI
2 Department of Pathology and Laboratory Medicine, University of Wisconsin, Madison, WI
3 Wisconsin National Primate Research Center, University of Wisconsin, Madison, WI; Department of Anatomy, University of Wisconsin, Madison, WI
* To whom correspondence should be addressed. E-mail: thomson{at}primate.wisc.edu.
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Abstract |
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Here we examine the ability of undifferentiated human ES cells to reprogram the nuclei of human ES cell-derived myeloid precursors following cell-cell fusion. Using an OP9 co-culture system, we produced CD45+CD33+myeloperoxidase(MPO)+ myeloid precursors from an Oct4-EGFP knock-in human ES cell line and demonstrated that Oct4-EGFP expression was extinguished in these precursors. Upon fusion with undifferentiated human ES cells, EGFP expression from the endogenous Oct4 promoter/regulatory region was re-established; ES cell-specific surface antigens and marker genes were expressed; and myeloid precursor-specific antigens were no longer detectable. When the hybrid cells were formed into embryoid bodies, up-regulation of genes characteristic of the three germ layers and extraembryonic tissues occurred, indicating that the hybrid cells had the potential to differentiate into multiple lineages. Interestingly, the hybrid cells were capable of re-differentiating into myeloid precursors with efficiency comparable to that of diploid human ES cells despite their near-tetraploid chromosome complement. These results indicate that human ES cells are capable of reprogramming nuclei from differentiated cells, and that human ES cell hybrid cells provide a new model system for studying the mechanisms of nuclear reprogramming.
Key Words. human embryonic stem cells, reprogramming, myeloid precursors, cell fusion, hybrid cells
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