Studies of the cell cycle of in vitro cultured skin fibroblasts in goats: work in progress

• Published in: Theriogenology 2003-03, Vol.59 (5), p.1277-1289
• Main Authors: Yu, Y.S., Sun, X.S., Jiang, H.N., Han, Y., Zhao, C.B., Tan, J.H.
• Format: Article
• Language: English
• Subjects: Animals; Apoptosis; Apoptosis - drug effects; Apoptosis - physiology; Bromodeoxyuridine - metabolism; Cell cycle; Cell Cycle - physiology; Cells, Cultured; Culture Media, Serum-Free; Enzyme Inhibitors - pharmacology; Female; Fibroblast; Fibroblasts - cytology; Fibroblasts - drug effects; Fibroblasts - metabolism; Flow Cytometry - veterinary; Goat; Goats; Kinetin; Olomoucine; Purines - pharmacology; Serum starvation; Skin - cytology; Skin - drug effects; Skin - metabolism
• ISSN: 0093-691X; 1879-3231
• DOI: 10.1016/S0093-691X(02)01193-7

The effect of serum starvation and olomoucine treatment on the cell cycle and apoptosis of goat skin fibroblasts cultured in vitro is reported in this paper. The cells were obtained from the ear of a female goat 1.5 years of age. Analysis of cell cycle distribution by fluorescence-activated cell sorting (FACS) showed that 3.4, 60.8 and 15.1% of normally cycling cells were at G1, G0 and S phase, respectively. Serum starvation for 1, 3 and 5 days arrested 70.1, 70.2 and 83.4% cells, respectively, at G0/G1 phase. Seventy-eight percent of confluent cells were at G0/G1 stage, but in contrast to the serum starved group, this high percentage of G0/G1 cells was mainly associated with G1 cells. Of cells not deprived of serum, 73.6% were arrested at G1/G0 when treated with 100 μM olomoucine for 9 h compared to 85.5% of cells that had been starved of serum for 2 days (co-inhibition) ( P0.05). When cells were co-inhibited, 4.1% of cells underwent apoptosis. In conclusion, although serum withdrawal for 5 days or more effectively arrested cells at G0/G1 stages, it increased apoptosis of cells significantly. However, co-inhibition by serum withdrawal and olomoucine treatment was found to be an appropriate treatment to obtain more healthy G0/G1 cells based on the low percentage of apoptotic cells after treatment.