Cell cycle analysis of bovine cultured somatic cells by flow cytometry
2003
Cheong, H.T. (Kangwon National Univ., Chunchon (Korea R.)) | Park, T.M. | Ikeda, K. | Takahashi, Y.
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 GO /G1, S and G2 /M. The majority was in GO /GI regardless of cell type and treatment. Serumstarved or confluent cultures contained higher percentages of cells in GO /G1 (89.5-95.4% ; P <0.05) . Percentages of cells in GO /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 GO /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 GO /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 GO /G1, and indicate that a more efficient synchronization of the cells in GO /G1 can be established by incubation for a limited time period after trypsinization of serum-starved or confluent cells.
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