Cell cycle analysis of in vitro cultured goral (Naemorhedus caudatus) adult skin fibroblasts.
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.
Hashem
Md Abul
MA
Department of Theriogenology and Biotechnology, College of Veterinary Medicine, Seoul National University, Seoul 151-742, South Korea.
Bhandari
Dilip P
DP
Kang
Sung Keun
SK
Lee
Byeong Chun
BC
Suk
Hwang Woo
HW
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*
D000975:Antioxidants; D016895:Culture Media, Serum-Free; D011480:Protease Inhibitors; D004121:Dimethyl Sulfoxide