Effects of serum deprivation and cycloheximide on cell cycle of low and high passage porcine fetal fibroblasts.
Arrest of cells in G0/G1 cell cycle phase is desired for nuclear transfer procedures. Serum starvation and cell cycle inhibitors are different ways to induce synchronization of the cell cycle. This study investigated the effects of serum starvation and cycloheximide (CHX) on the cell cycle of low (5th) and high (15th) passages fetal porcine fibroblasts. Cell cycle phases were determined using fluorescent activated cell sorting. Fifth passage fibroblast cultures had higher (p < 0.05) proportion of cells in G0/G1 only after 72 h of serum starvation (77.60 +/- 0.65) when compared with non-starved cells (71.44 +/- 1.88). Serum starvation for all periods tested induced an increase (p < 0.05) on proportion of cells in G0/G1 on the 15th passage. No significant differences were observed on the 5th passage cultures exposed to CHX, although, on the 15th passage an increase on proportion of cells was observed after all periods of exposure (p < 0.05). These data indicates that high passage cells in vitro are more susceptible to serum starvation and CHX G0/G1 synchronization.
Goissis
M D
MD
Laboratory of In vitro Fertilization, Cloning and Animal Transgenesis, Department of Animal Reproduction, Faculty of Veterinary Medicine and Animal Sciences, University of São Paulo, São Paulo, Brazil.
Caetano
H V A
HV
Marques
M G
MG
de Barros
F R O
FR
Feitosa
W B
WB
Milazzotto
M P
MP
Binelli
M
M
Assumpção
M E O A
ME
Visintin
J A
JA
D000818:Animals; D002453:Cell Cycle / Q000187:drug effects* / Q000502:physiology; D002455:Cell Division / Q000502:physiology; D002469:Cell Separation / Q000662:veterinary; D002470:Cell Survival; D002478:Cells, Cultured; D016895:Culture Media, Serum-Free / Q000494:pharmacology*; D003513:Cycloheximide / Q000494:pharmacology*; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D005434:Flow Cytometry / Q000662:veterinary; D016193:G1 Phase; D016192:Resting Phase, Cell Cycle; D013552:Swine / Q000196:embryology*
D016895:Culture Media, Serum-Free; D003513:Cycloheximide