Antiapoptotic and embryotrophic effects of alpha-tocopherol and L-ascorbic acid on porcine embryos derived from in vitro fertilization and somatic cell nuclear transfer.
The objective was to determine optimal concentrations of alpha-tocopherol and l-ascorbic acid for development of porcine embryos derived from in vitro-fertilization (IVF) or somatic cell nuclear transfer (SCNT). The frequency of blastocyst formation in IVF embryos was 17.6, 28.6, 32.4 and 21.4% for control, 50, 100 and 200microM alpha-tocopherol, respectively, whereas in SCNT embryos, the frequency was 12.8, 19.0, 24.8 and 17.7% for corresponding concentrations of alpha-tocopherol. For both IVF and SCNT embryos, there were significantly more cells in blastocysts and the embryos had greater developmental competence when the embryo culture medium was supplemented with 100microM alpha-tocopherol. Although either alpha-tocopherol or l-ascorbic acid reduced the proportion of apoptotic cells in blastocysts, in combination they resulted in rates of apoptosis that were similar to the control group. For IVF embryos, the apoptotic index was 0.09 and 0.11 for alpha-tocopherol or l-ascorbic acid at a concentration of 100microM, respectively. Conversely, when these antioxidants were supplemented together, the apoptotic index increased significantly and was similar to the control group (i.e., 0.17 and 0.21 for combined and control group). For SCNT embryos, the apoptotic index was 0.10 for 100microM for both alpha-tocopherol and l-ascorbic acid, whereas the index was 0.23 and 0.17 for control and combined group. In conclusion, we recommend supplementing porcine embryo culture medium with 100microM alpha-tocopherol or 100microM l-ascorbic (as a second choice).
Jeong
Yeon Woo
YW
Department of Theriogenology and Biotechnology, College of Veterinary Medicine, Seoul National University, 56-1 Sillim-dong, Gwanak-gu, Seoul 151-742, Korea.
Park
Sun Woo
SW
Hossein
Mohammad Shamim
MS
Kim
Sue
S
Kim
Ji Hye
JH
Lee
So Hyun
SH
Kang
Sung Keun
SK
Lee
Byeong Chun
BC
Hwang
Woo Suk
WS
D000818:Animals; D000975:Antioxidants / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects*; D001205:Ascorbic Acid / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D046149:Embryo Culture Techniques / Q000379:methods / Q000662:veterinary; D047108:Embryonic Development / Q000187:drug effects*; D005307:Fertilization in Vitro / Q000662:veterinary; D020287:In Situ Nick-End Labeling; D053652:Nuclear Transfer Techniques / Q000662:veterinary; D013552:Swine / Q000196:embryology*; D024502:alpha-Tocopherol / Q000494:pharmacology*
D000975:Antioxidants; D024502:alpha-Tocopherol; D001205:Ascorbic Acid