Abstract
Here we describe a lineage reprogramming system consisting of a B cell line with an estradiol-inducible form of C/EBPalpha where cells can be converted into macrophage-like cells at 100% efficiency within 2 to 3 days. The reprogrammed cells are larger, contain altered organelle and cytoskeletal structures, are phagocytic, and exhibit an inflammatory response. Time-lapse experiments showed that the cells acquire a macrophage morphology and increased migratory activity as early as 10 hr. During induction, thousands of genes become up- or downregulated, including several dozen transcription and chromatin-remodeling factors. Time-limited exposure of cells to the inducer showed that the reprogrammed cells become transgene independent within 1 to 2 days. The reprogramming can be inhibited, at least partially, by perturbation experiments with B cell and macrophage transcription factors. The tightness, robustness, and speed of the system described make it a versatile tool to study biochemical and biological aspects of lineage reprogramming.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Antigens, Differentiation / metabolism
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Basic-Leucine Zipper Transcription Factors / genetics
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Basic-Leucine Zipper Transcription Factors / metabolism*
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Cell Line, Transformed
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Cell Movement / genetics
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Cell Movement / immunology
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Cell Transdifferentiation*
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DNA-Binding Proteins / genetics
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DNA-Binding Proteins / metabolism
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Estradiol / metabolism
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Estrogen Receptor alpha / genetics
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Gene Expression Profiling
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Gene Expression Regulation, Developmental / immunology
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Inflammation Mediators / metabolism
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Macrophages / cytology
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Macrophages / immunology
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Macrophages / metabolism*
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Mice
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Precursor Cells, B-Lymphoid / cytology
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Precursor Cells, B-Lymphoid / immunology
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Precursor Cells, B-Lymphoid / metabolism*
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Recombinant Fusion Proteins / genetics
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Recombinant Fusion Proteins / metabolism*
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Transcription Factor AP-2
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Transcriptional Activation
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Transduction, Genetic
Substances
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Antigens, Differentiation
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Basic-Leucine Zipper Transcription Factors
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DNA-Binding Proteins
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Estrogen Receptor alpha
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Inflammation Mediators
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Recombinant Fusion Proteins
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Tfap2a protein, mouse
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Transcription Factor AP-2
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Estradiol