Reprogramming fibroblasts to express T-cell functions using cell extracts.
We demonstrate here the functional reprogramming of a somatic cell using a nuclear and cytoplasmic extract derived from another somatic cell type. Reprogramming of 293T fibroblasts in an extract from primary human T cells or from a transformed T-cell line is evidenced by nuclear uptake and assembly of transcription factors, induction of activity of a chromatin remodeling complex, histone acetylation, and activation of lymphoid cell specific genes. Reprogrammed cells express T cell specific receptors and assemble the interleukin-2 receptor in response to T cell receptor CD3 (TCR CD3) complex stimulation. Reprogrammed primary skin fibroblasts also express T cell specific antigens. After exposure to a neuronal precursor extract, 293T fibroblasts express a neurofilament protein and extend neurite-like outgrowths. In vitro reprogramming of differentiated somatic cells creates possibilities for producing isogenic replacement cells for therapeutic applications.
Håkelien
Anne-Mari
AM
Institute of Medical Biochemistry, P.O. Box 1112, Blindern, University of Oslo, Oslo 0317, Norway.
Landsverk
Helga B
HB
Robl
James M
JM
Skålhegg
Bjørn S
BS
Collas
Philippe
P
D001709:Biotechnology / Q000379:methods; D017252:CD3 Complex / Q000378:metabolism; D002454:Cell Differentiation; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism; D002843:Chromatin / Q000378:metabolism; D003593:Cytoplasm / Q000378:metabolism; D005347:Fibroblasts / Q000378:metabolism*; D005821:Genetic Techniques; D006657:Histones / Q000378:metabolism; D006801:Humans; D007376:Interleukin-2 / Q000378:metabolism; D019169:Jurkat Cells; D008856:Microscopy, Fluorescence; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013601:T-Lymphocytes / Q000502:physiology*; D013997:Time Factors; D014158:Transcription, Genetic
D017252:CD3 Complex; D002843:Chromatin; D006657:Histones; D007376:Interleukin-2; D012333:RNA, Messenger