Transient alteration of cell fate using a nuclear and cytoplasmic extract of an insulinoma cell line.
We report a transient modulation of cell fate in fibroblasts briefly exposed to an extract derived from the rat insulin-producing beta cell line, INS-1E. Primary fetal rat fibroblasts were reversibly permeabilized with Streptolysin O, incubated for 1h in a 15,000g INS-1E nuclear and cytoplasmic extract, resealed, and cultured. A first marker of change in cell fate was a reduction of cell and nuclear size within days of exposure to extract such that in some instances the fibroblasts resembled INS-1E cells. Second, two beta cell transcripts, Pdx-1 and insulin, were detected in the fibroblasts for up to 4 weeks. Third, (pro)insulin labeling was detected in 5-30% of the cells for a period of 8-14 days after incubation in extract. These phenotypes were absent from fibroblasts exposed to heat-treated INS-1E extracts, a human fibroblast cell line-derived extract or buffer. The results indicate that the extract of an insulinoma-derived cell line can promote at least a transient modification of cell fate towards a beta cell phenotype in non-beta cells. Because they are easily accessible, cell extracts may represent a practical source of material for investigating the mechanisms of alteration of a nuclear and cellular program.
Håkelien
Anne-Mari
AM
Institute of Medical Biochemistry, University of Oslo, Oslo 0317, Norway.
Gaustad
Kristine G
KG
Collas
Philippe
P
D000818:Animals; D001426:Bacterial Proteins; D018929:Cell Culture Techniques / Q000379:methods*; D002460:Cell Line; D019070:Cell Lineage; D002467:Cell Nucleus / Q000378:metabolism*; D002478:Cells, Cultured; D003593:Cytoplasm / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D005347:Fibroblasts / Q000378:metabolism; D018398:Homeodomain Proteins; D007328:Insulin / Q000378:metabolism; D007340:Insulinoma / Q000378:metabolism*; D008856:Microscopy, Fluorescence; D010641:Phenotype; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013301:Streptolysins / Q000494:pharmacology; D013997:Time Factors; D015534:Trans-Activators / Q000378:metabolism; D014158:Transcription, Genetic
D001426:Bacterial Proteins; D018398:Homeodomain Proteins; D007328:Insulin; D012333:RNA, Messenger; D013301:Streptolysins; D015534:Trans-Activators; C084010:pancreatic and duodenal homeobox 1 protein; C025964:streptolysin O