Cell cycle analysis of bovine cultured somatic cells by flow cytometry

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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.
北海道大学の論文
2003-08-29

Cell cycle analysis of bovine cultured somatic cells by flow cytometry

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