DNA methylation in bovine adult and fetal fibroblast cells.
The aim of this project was to develop a simple screening tool to measure the DNA methylation of fibroblast cells, and to determine if differences in DNA methylation could be detected in adult and fetal fibroblast cells after serum starvation (SS). Four adult and four fetal tissue explants were collected to produce presumptive fibroblast cell cultures for this experiment. All cell lines underwent three repetitions of serum starvation for 0 (control), 2, 5, or 7 days. The DNA was extracted from the cells and analyzed for DNA methylation content using methylation sensitive restriction enzyme digestion, gel electrophoresis and image analysis. There was no difference (p = 0.11) between the DNA methylation of the adult and fetal nonclonal cell lines. A cubic trend (p = 0.09) of increased DNA methylation at 2 days of serum starvation followed by periods of decreasing DNA methylation at 5 and 7 days were observed for the adult nonclonal cell lines. A significant interaction (p = 0.03) was observed between fetal cell line and day. This simple, rapid DNA methylation assay may be beneficial when evaluating cells' DNA methylation content.
Jones
Karen L
KL
Animal Science, Food and Nutrition, Southern Illinois University-Carbondale, Carbondale, Illinois, USA. kljones@siu.edu
Tarochione-Utt
Kelly D
KD
D000328:Adult; D000818:Animals; D002417:Cattle; D002478:Cells, Cultured; D016895:Culture Media, Serum-Free; D019175:DNA Methylation; D005260:Female; D005333:Fetus / Q000166:cytology; D005347:Fibroblasts / Q000378:metabolism*; D006801:Humans; D011247:Pregnancy
D016895:Culture Media, Serum-Free