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Synchronizing cell cycle of goat fibroblasts by serum starvation causes apoptosis.
MedLine Citation:
PMID:  19788523     Owner:  NLM     Status:  In-Data-Review    
Abstract/OtherAbstract:
Cell cycle stage and synchronization of donor cells are important factors influencing the success of somatic cell nuclear transfer. This study examined whether serum starvation has any effect on specific cell death. We also studied the effects of serum starvation, culture to confluence, and full confluency (confluent + 72 h) on cell cycle characteristics and apoptosis of goat dermal fibroblast cells. The cells were obtained from the ear of a 1.5-year-old female goat. The following experimental groups were analysed for fibroblast cells: (i) normally growing, (ii) confluent, (iii) full confluency, (iv) cells starved for 48 h and (v) cells starved for 72 h. Analysis of cell cycle distribution by flow cytometry showed that 4.56 and 51.88% of normal cycling cells were at the G0 and G1 phases respectively. In the confluent group, 80% of the cells were arrested in the G0/G1 phase. Serum starvation for 48 and 72 h arrested 84.78% and 90.1% cells at the G0/G1 phase respectively which showed a significant difference when compared with the control group (p < 0.05). Double staining by PI and FITC distinguishes G0 phase from G1 phase. In the full confluency group, 91.53% of 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. Under normal culture conditions, 6.39% of cells underwent early apoptosis. In the confluent group 8.93% of cells showed early apoptosis. Serum starvation for 48 and 72 h caused early apoptosis in 8.91 and 39.83% of the cells respectively. Full confluency treatment did not increase the number of apoptotic cells significantly (8.67%). After 72 h, serum starvation significantly increased early apoptosis (p < 0.05). In conclusion, the use of full confluency is suitable for cell cycle synchronization because it arrests cells at the G0/G1 phase and also induces less apoptosis in comparison with the serum starvation group.
Authors:
A Dalman; P Eftekhari-Yazdi; Mr Valojerdi; A Shahverdi; H Gourabi; E Janzamin; R Fakheri; F Sadeghian; F Hasani
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Reproduction in domestic animals = Zuchthygiene     Volume:  45     ISSN:  1439-0531     ISO Abbreviation:  Reprod. Domest. Anim.     Publication Date:  2010 Oct 
Date Detail:
Created Date:  2011-04-04     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9015668     Medline TA:  Reprod Domest Anim     Country:  Germany    
Other Details:
Languages:  eng     Pagination:  e46-53     Citation Subset:  IM    
Copyright Information:
© 2009 Blackwell Verlag GmbH.
Affiliation:
Embryology Department, Cell Sciences Research Center, Royan Institute, ACECR, Tehran, Iran Genetic Department, Cell Sciences Research Center, Royan Institute, ACECR, Tehran, Iran Stem Cell Department, Cell Sciences Research Center, Royan Institute, ACECR, Tehran, Iran Research and Clinical Center for Infertility, Shahid Sadoughi University of Medical Science, Yazd, Iran.
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