A robust and highly efficient immune cell reprogramming system.
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.
Bussmann
Lars H
LH
Center for Genomic Regulation and Universitat Pompeu Fabra, Barcelona, Spain.
Schubert
Alexis
A
Vu Manh
Thien Phong
TP
De Andres
Luisa
L
Desbordes
Sabrina C
SC
Parra
Maribel
M
Zimmermann
Timo
T
Rapino
Francesca
F
Rodriguez-Ubreva
Javier
J
Ballestar
Esteban
E
Graf
Thomas
T
D000818:Animals; D000943:Antigens, Differentiation / Q000378:metabolism; D050976:Basic-Leucine Zipper Transcription Factors / Q000235:genetics / Q000378:metabolism*; D002461:Cell Line, Transformed; D002465:Cell Movement / Q000235:genetics / Q000276:immunology; D054338:Cell Transdifferentiation; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D004958:Estradiol / Q000378:metabolism; D047628:Estrogen Receptor alpha / Q000235:genetics; D020869:Gene Expression Profiling; D018507:Gene Expression Regulation, Developmental / Q000276:immunology; D018836:Inflammation Mediators / Q000378:metabolism; D008264:Macrophages / Q000166:cytology / Q000276:immunology / Q000378:metabolism*; D051379:Mice; D054448:Precursor Cells, B-Lymphoid / Q000166:cytology / Q000276:immunology / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism*; D050656:Transcription Factor AP-2; D015533:Transcriptional Activation; D014161:Transduction, Genetic
D000943:Antigens, Differentiation; D050976:Basic-Leucine Zipper Transcription Factors; D004268:DNA-Binding Proteins; D047628:Estrogen Receptor alpha; D018836:Inflammation Mediators; D011993:Recombinant Fusion Proteins; C493668:Tfap2a protein, mouse; D050656:Transcription Factor AP-2; D004958:Estradiol