Abstract
The objective of this study was to investigate the effects of cell status of BOEC on development of bovine IVM/IVF embryos and gene expression in BOEC before or after culturing of embryos. The developmental rates beyond morula stage in the BOEC co-culture group was significantly higher than in the control group (p<0.05). In particular, blastocyst production in the BOEC co-culture group (28.3%) was dramatically increased compared with the control group (7.2%). In the in vitro development of bovine IVM/IVF embryos according to cell status, the developmental rates beyond morula stage in the primary culture cell (PCC) co-culture group were the highest of all experimental groups. Expression of genes related to growth (TGF-β, EGF and IGFBP), apoptosis (Bax, Caspase-3 and p53) and antioxidation (CuZnSOD, MnSOD, Catalase and GPx) in different status cells of BOEC for embryo culture was detected by RT-PCR. While EGF gene was detected in isolated fresh cells (IFC) and PCC, TGF-β and IGFBP were found in IFC or PCC after use in the embryo culture, respectively. Caspase-3 and Bax genes were detected in all experimental groups regardless of whether the BOEC was used or not used in the embryo culture. However, p53 gene was found in IFC of both conditions for embryo culture and in frozen/thawed culture cells (FPCC) after use in the embryo culture. Although antioxidant genes examined were detected in all experimental groups before using for the embryo culture, these genes were not detected after use. This study indicated that the BOEC co-culture system used for in vitro culture of bovine IVF embryos can increase the developmental rates, and cell generations and status of BOEC might affect the in vitro development of bovine embryos. The BOEC monolayer used in the embryo culture did not express the growth factors (TGF-β and EGF) and enzymatic antioxidant genes, thereby improving embryo development in vitro.
Highlights
Since the birth of the first calf derived from in vitro fertilization (Brackett et al, 1982), considerable progress has been made in the development of techniques for in vitro production (IVP) of bovine embryos for both research and commercial purposes
The goal of the present study was to investigate the factors affecting increased in vitro developmental rates when Hanwoo IVM/IVF embryos were co-cultured with bovine oviduct epithelial cell (BOEC)
The deleterious conditions for in vitro culture of implantation embryos are the exposure of ROS by high oxygen tension, heavy metal ions, unknown embryotoxic substances and others
Summary
Since the birth of the first calf derived from in vitro fertilization (Brackett et al, 1982), considerable progress has been made in the development of techniques for in vitro production (IVP) of bovine embryos for both research and commercial purposes. The co-culture system with somatic cells is an effective and useful method for in vitro culture of bovine IVF embryos, affecting the success of IVP systems which may stimulate embryo development through amelioration of embryotoxic substance or secretion of embryotropic factor (Carolan et al, 1995; Donnay et al, 1997). The goal of the present study was to investigate the factors affecting increased in vitro developmental rates when Hanwoo IVM/IVF embryos were co-cultured with BOEC. The present study was designed to determine 1) the effects of the BOEC co-culture system on in vitro development rate of bovine IVM/IVF embryos, 2) the effect of status of BOEC on in vitro development of bovine adjusted to 3×106 cell/ml with DMEM containing 10% FBS.
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