title: Cell cycle analysis of bovine cultured somatic cells by flow cytometry.
abstract: This study was undertaken to examine the cell cycle characteristics of bovine fetal and adult somatic cells (fetal fibroblasts, adult skin and muscle cells, and cumulus cells) after culture under a variety of conditions; 1) growth to 60-70% confluency (cycling), 2) serum starvation, 3) culture to confluency. Cell -cycle phases were determined by flow cytometry with propidium iodide staining enabling the calculation of percentages of cells in G0 /G1, S and G2 /M. The majority was in G0/G1 regardless of cell type and treatment. Serum-starved or confluent cultures contained higher percentages of cells in G0/G1 (89.5-95.4%; P < 0.05). Percentages of cells in G0/G1 increased as cell size decreased regardless of the cell type and treatment. In the serum-starved and confluent cultures, about 98% of small cells were in G0/G1 . Serum-starved cultures contained higher percentages of small cells (38.5-66.9%) than cycling and confluent cultures regardless of cell type (P < 0.05) . After trypsinization of fetal fibroblasts and adult skin cells that were serum-starved and cultured to confluency, the percentages of cells in G0/G1 increased (P < 0.05) on incubation for 1.5 (95.7-99.5%) or 3 hr (95.9-98.6%). These results verify that serum starvation and culture to confluency are efficient means of synchronizing bovine somatic cells in G0/G1, and indicate that a more efficient synchronization of the cells in G0/G1 can be established by incubation for a limited time period after trypsinization of serum-starved or confluent cells.
authors: Department of Veterinary Medicine, College of Animal Resource Science, Kangwon National University, Chuncheon 200-701, Korea.
authors: Hee-Tae
authors: HT
authors: Cheong
authors: Tae-Mook
authors: TM
authors: Park
authors: Koji
authors: K
authors: Ikeda
authors: Yoshiyuki
authors: Y
authors: Takahashi
mesh_terms: D000818:Animals; D002417:Cattle; D002453:Cell Cycle; D002478:Cells, Cultured; D016895:Culture Media, Serum-Free; D005260:Female; D005333:Fetus / Q000166:cytology; D005347:Fibroblasts / Q000166:cytology; D005434:Flow Cytometry / Q000662:veterinary*; D008297:Male; D032342:Muscle Cells / Q000166:cytology; D006080:Ovarian Follicle / Q000166:cytology; D012867:Skin / Q000166:cytology
chemical_list: D016895:Culture Media, Serum-Free