title: Effect of Acteoside as a Cell Protector to Produce a Cloned Dog.
abstract: Somatic cell nuclear transfer (SCNT) is a well-known laboratory technique. The principle of the SCNT involves the reprogramming a somatic nucleus by injecting a somatic cell into a recipient oocyte whose nucleus has been removed. Therefore, the nucleus donor cells are considered as a crucial factor in SCNT. Cell cycle synchronization of nucleus donor cells at G0/G1 stage can be induced by contact inhibition or serum starvation. In this study, acteoside, a phenylpropanoid glycoside compound, was investigated to determine whether it is applicable for inducing cell cycle synchronization, cytoprotection, and improving SCNT efficiency in canine fetal fibroblasts. Primary canine fetal fibroblasts were treated with acteoside (10, 30, 50 μM) for various time periods (24, 48 and 72 hours). Cell cycle synchronization at G0/G1 stage did not differ significantly with the method of induction: acteoside treatment, contact inhibition or serum starvation. However, of these three treatments, serum starvation resulted in significantly increased level of reactive oxygen species (ROS) (99.5 ± 0.3%) and apoptosis. The results also revealed that acteoside reduced ROS and apoptosis processes including necrosis in canine fetal fibroblasts, and improved the cell survival. Canine fetal fibroblasts treated with acteoside were successfully arrested at the G0/G1 stage. Moreover, the reconstructed embryos using nucleus donor cells treated with acteoside produced a healthy cloned dog, but not the embryos produced using nucleus donor cells subjected to contact inhibition. In conclusion, acteoside induced cell cycle synchronization of nucleus donor cells would be an alternative method to improve the efficiency of canine SCNT because of its cytoprotective effects.
authors.affiliation: Division of Animal & Dairy Science, College of Agriculture and Life Science, Chungnam National University, Daejeon, Republic of Korea.
authors.forename: Ji Hye
authors.initials: JH
authors.lastname: Lee
authors.affiliation: Division of Animal & Dairy Science, College of Agriculture and Life Science, Chungnam National University, Daejeon, Republic of Korea.
authors.forename: Ju Lan
authors.initials: JL
authors.lastname: Chun
authors.affiliation: Division of Animal & Dairy Science, College of Agriculture and Life Science, Chungnam National University, Daejeon, Republic of Korea.
authors.forename: Keun Jung
authors.initials: KJ
authors.lastname: Kim
authors.affiliation: Division of Animal & Dairy Science, College of Agriculture and Life Science, Chungnam National University, Daejeon, Republic of Korea.
authors.forename: Eun Young
authors.initials: EY
authors.lastname: Kim
authors.affiliation: Division of Animal & Dairy Science, College of Agriculture and Life Science, Chungnam National University, Daejeon, Republic of Korea.
authors.forename: Dong-Hee
authors.initials: DH
authors.lastname: Kim
authors.affiliation: Division of Animal & Dairy Science, College of Agriculture and Life Science, Chungnam National University, Daejeon, Republic of Korea.
authors.forename: Bo Myeong
authors.initials: BM
authors.lastname: Lee
authors.affiliation: Division of Animal & Dairy Science, College of Agriculture and Life Science, Chungnam National University, Daejeon, Republic of Korea.
authors.forename: Kil Woo
authors.initials: KW
authors.lastname: Han
authors.affiliation: Division of Animal & Dairy Science, College of Agriculture and Life Science, Chungnam National University, Daejeon, Republic of Korea.
authors.forename: Kang-Sun
authors.initials: KS
authors.lastname: Park
authors.affiliation: Department of Biology Education, College of Education, Chonnam National University, Gwangju, Republic of Korea.
authors.forename: Kyung-Bon
authors.initials: KB
authors.lastname: Lee
authors.affiliation: Division of Animal & Dairy Science, College of Agriculture and Life Science, Chungnam National University, Daejeon, Republic of Korea.
authors.forename: Min Kyu
authors.initials: MK
authors.lastname: Kim
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D002467:Cell Nucleus / Q000187:drug effects* / Q000502:physiology; D002470:Cell Survival / Q000187:drug effects; D019976:Cloning, Organism / Q000379:methods; D003260:Contact Inhibition / Q000502:physiology; D016895:Culture Media, Serum-Free / Q000494:pharmacology; D019610:Cytoprotection / Q000502:physiology*; D004285:Dogs; D004624:Embryo Transfer; D005260:Female; D005333:Fetus; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects* / Q000502:physiology; D016193:G1 Phase / Q000187:drug effects*; D005960:Glucosides / Q000494:pharmacology*; D053652:Nuclear Transfer Techniques; D010636:Phenols / Q000494:pharmacology*; D017382:Reactive Oxygen Species / Q000378:metabolism; D059808:Polyphenols
chemical_list: D016895:Culture Media, Serum-Free; D005960:Glucosides; D010636:Phenols; D017382:Reactive Oxygen Species; C058956:acteoside; D059808:Polyphenols