Flow cytometric cell-cycle analysis of cultured fibroblasts from the giant panda, Ailuropoda melanoleuca L.
In animal cloning, it is generally believed that the inactive diploid G(0)or G(1)stage of the cell cycle is beneficial to initiate cell-cycle coordination and reprogramming following transfer of the donor nucleus. Previous experiments have demonstrated that serum starvation results in quiescent cell stage. Some experiments show that the majority of cells in a fully confluent cell culture are also in an inactive G(1)stage. In order to provide more G(0)/G(1)stage cells for giant panda cloning, we carried out a flow cytometric analysis of the cell cycle of fibroblasts from the abdominal muscle of a giant panda at different passage numbers under different growth conditions, and after different periods of serum starvation. The percentage of G(0)+G(1)stage cells differed significantly under different growth conditions. Serum starvation effectively increased the percentage of G(0)+G(1)stage cells, and the cell cycle characteristics following serum starvation for varying periods of time differed with this and the initial confluency of the cultures. The data should help in choosing the optimal stage for preparing donor cells as well as increasing the potential cloning efficiency in our study of giant panda cloning.
Han
Zhi-Ming
ZM
State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100080, People's Republic of China.
Chen
Da-Yuan
DY
Li
Jin-Song
JS
Sun
Qing-Yuan
QY
Wang
Peng-Yan
PY
Du
Jun
J
Zhang
He-Min
HM
D000818:Animals; D002453:Cell Cycle / Q000187:drug effects; D002478:Cells, Cultured; D016895:Culture Media, Serum-Free / Q000494:pharmacology; D005260:Female; D005347:Fibroblasts / Q000166:cytology* / Q000187:drug effects; D005434:Flow Cytometry / Q000379:methods*; D001503:Ursidae
D016895:Culture Media, Serum-Free