title: Cell cycle analysis of in vitro cultured goral (Naemorhedus caudatus) adult skin fibroblasts.
abstract: The present study was undertaken to examine cell cycle characteristics of endangered Goral (CITES Appendix I) adult skin fibroblasts. Seven experiments were performed, each with a one-way completely randomized design involving three replicates. Least significant difference (LSD) was used to determine variation among treatment groups. Experiment I focused on the effects of cycling, serum-starved, and fully confluent stages of Goral cells. In Experiments II and III, the effects of different antioxidants like beta-mercaptoethanol (beta-ME, 10 microM), cysteine (2 mM), and glutathione (2 mM) were examined after cells were fully confluent without serum starvation for 24 h and 4 h, respectively. In Experiments IV and V, three protease inhibitors, namely 6-dimethylaminopurine (6-DMAP, 2 mM), cycloheximide (7.5 microg/ml) and cytochalasin B (7.5 microg/ml), were used as in Experiment II. In Experiments VI and VII, the effect of different levels of dimethylsulphoxide (DMSO) at 0%, 0.5%, 1.0% and 2.5% were tested by flow cytometry (FACS). In Experiment I, 68.7% of Goral skin fibroblasts reached the G(0)/G(1) stage (2C DNA content) when subjected to the serum-starved medium, which was more than the cycling (64.9%) and fully confluent groups (61.0%) (P > 0.05). Among the chemically treated group, beta-ME, cysteine and DMSO showed better results for synchronization of G(0) + G(1) phases than cycling, serum-starved and fully confluent groups. It can thus be concluded that beta-ME, cysteine and DMSO at certain concentrations can synchronize the cell cycle effectively, which could have a positive impact on somatic cell nuclear transfer in the goral.
authors.affiliation: Department of Theriogenology and Biotechnology, College of Veterinary Medicine, Seoul National University, Seoul 151-742, South Korea.
authors.forename: Md Abul
authors.initials: MA
authors.lastname: Hashem
authors.forename: Dilip P
authors.initials: DP
authors.lastname: Bhandari
authors.forename: Sung Keun
authors.initials: SK
authors.lastname: Kang
authors.forename: Byeong Chun
authors.initials: BC
authors.lastname: Lee
authors.forename: Hwang Woo
authors.initials: HW
authors.lastname: Suk
mesh_terms: D000818:Animals; D000975:Antioxidants / Q000494:pharmacology; D002453:Cell Cycle / Q000187:drug effects; D002478:Cells, Cultured; D016895:Culture Media, Serum-Free; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D005260:Female; D005347:Fibroblasts / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D011480:Protease Inhibitors / Q000494:pharmacology; D012418:Ruminants; D012867:Skin / Q000166:cytology*
chemical_list: D000975:Antioxidants; D016895:Culture Media, Serum-Free; D011480:Protease Inhibitors; D004121:Dimethyl Sulfoxide