Embryotrophic effects of ethylenediaminetetraacetic acid and hemoglobin on in vitro porcine embryos development.
This study investigated the embryotrophic effects of ethylenediaminetetraacetic acid (EDTA) and hemoglobin (Hb) on porcine preimplantation embryo development. Porcine embryos produced by in vitro maturation/fertilization were cultured for 6 days in modified North Carolina State University-23 medium (mNCSU-23) supplemented with EDTA and/or Hb. In Exp. 1, culturing porcine zygotes with 100 microM EDTA significantly increased cleavage frequencies (85.3%) at 48 h post insemination and the number of inner cell mass (ICM) (9.6+/-5.5) compared to the control (7.0+/-2.8). However, 100 microM EDTA did not improve blastocyst formation compared to 0, 1 or 10 microM EDTA. In Exp. 2, in vitro fertilized oocytes were cultured with 0, 1 or 10 microg/ml Hb. Culturing with Hb did not promote porcine embryo development, but significantly increased the cell numbers of blastocysts in 1 microg/ml Hb compared to 0 or 10 microg/ml Hb. In Exp. 3, culturing embryos with 100 microM EDTA+1 microg/ml Hb significantly improved frequencies of cleavage, blastocyst formation, and total cell numbers in blastocysts compared to the control. Moreover, 100 microM EDTA, 1 microg/ml Hb and their combination reduced reactive oxygen species (ROS) accumulation and decreased the incidence of apoptosis. In conclusion, the present study clearly demonstrated that the combining treatment of EDTA and Hb improved IVF porcine embryo development.
Kim
Ji Hye
JH
Department of Theriogenology and Biotechnology, College of Veterinary Medicine, Seoul National University, Seoul 151-742, South Korea.
Lee
So Hyun
SH
Kim
Sue
S
Jeong
Yeon Woo
YW
Koo
Ok Jae
OJ
Hashem
M D Abul
MD
Park
Seon Mi
SM
Lee
Eu Gine
EG
Hossein
M S
MS
Kang
Sung Keun
SK
Lee
Byeong Chun
BC
Hwang
Woo Suk
WS
D000818:Animals; D004305:Dose-Response Relationship, Drug; D004492:Edetic Acid / Q000494:pharmacology*; D046149:Embryo Culture Techniques / Q000379:methods / Q000662:veterinary*; D047108:Embryonic Development / Q000187:drug effects*; D005307:Fertilization in Vitro / Q000379:methods / Q000662:veterinary*; D006454:Hemoglobins / Q000494:pharmacology*; D020287:In Situ Nick-End Labeling; D017382:Reactive Oxygen Species / Q000032:analysis / Q000378:metabolism; D013552:Swine / Q000196:embryology*
D006454:Hemoglobins; D017382:Reactive Oxygen Species; D004492:Edetic Acid