title: In vitro reprogramming of nuclei and cells.
abstract: Directly turning a somatic cell type into another would be beneficial for producing replacement cells for therapeutic purposes. To this end, novel cell reprogramming strategies are being developed. We describe here methods for functionally reprogramming a somatic cell using an extract derived from another somatic cell type. The procedure involves reversible permeabilization of 293T fibroblasts, incubation of the permeabilized cells in a nuclear and cytoplasmic extract of T-cells, resealing of the "reprogrammed" cells, and culture for assessment of reprogramming. Reprogramming has been evidenced by nuclear uptake and assembly of transcription factors, induction of activity of a chromatin remodeling complex, changes in chromatin composition, activation of lymphoid cell-specific genes, and expression of T-cell-specific surface molecules. The system is likely to constitute a powerful tool to examine the processes of nuclear reprogramming, at least as they occur in vitro.
authors.affiliation: Institute of Medical Biochemistry, University of Oslo, Oslo, Norway.
authors.forename: Anne-Mari
authors.initials: AM
authors.lastname: Håkelien
authors.forename: Thomas
authors.initials: T
authors.lastname: Küntziger
authors.forename: Kristine G
authors.initials: KG
authors.lastname: Gaustad
authors.forename: Anne
authors.initials: A
authors.lastname: Marstad
authors.forename: Philippe
authors.initials: P
authors.lastname: Collas
mesh_terms: D001426:Bacterial Proteins / Q000493:pharmacokinetics; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation; D002457:Cell Extracts / Q000235:genetics; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D002467:Cell Nucleus / Q000235:genetics / Q000378:metabolism*; D005347:Fibroblasts / Q000502:physiology; D006801:Humans; D010539:Permeability; D013301:Streptolysins / Q000493:pharmacokinetics
chemical_list: D001426:Bacterial Proteins; D002457:Cell Extracts; D013301:Streptolysins; C025964:streptolysin O