Fetal Bovine Serum (FBS) plays a crucial role in animal cell culture; however, the increasing number of bovine fetuses used and sacrificed solely for FBS collection has raised ethical concerns globally. The welfare of fetuses during FBS blood collection has become a key focus of debate among animal welfare and ethics organizations worldwide. Previous studies indicate that heat-inactivated coelomic fluid (HI-CF) from the earthworm Perionyx excavatus may serve as a viable FBS alternative in adherent cell cultures. This study evaluates the potential of HI-CF as an FBS substitute during the in vitro maturation (IVM) stage of bovine embryo culture, with a focus on improving developmental rate through antioxidation effects. In this study, 2% HI-CF was incorporated into IVM media, assessing its impact on cell growth, differentiation, and the expression of genes related to antioxidation. The group of 2% of HI-CF exhibited a trend toward increased cleavage and blastocyst development rates compared to the control group. Although antioxidant genes such as NRF2 and GSR showed no statistically significant differences between the control and treatment groups, a trend toward increased expression was observed. Conversely, GPX1 displayed a trend of decreased expression. Notably, IGF1 and NQO1 were significant upregulated (p < 0.05) in the 2% HI-CF group. Additionally, oocytes stained with H2DCFDA showed a significantly reduced ROS levels (p < 0.05) in the 2% HI-CF group compared with controls. These findings suggest that HI-CF's antioxidative effects support enhanced cell growth and blastocyst development rate, surpassing those observed with FBS. Consequently, HI-CF shows promise as an effective alternative to FBS in vitro maturation of bovine oocytes.

To investigated the mechanism, induced pluripotent stem cells(iPSC) is important for clinical application and stem cell research. It is well known that hMAGEA2 expression pattern and effect on differentiation in embryonic stem cell but their specific role in iPS cells are unclear. The present study was schemed to understand the function of hMAGEA2 gene in iPS cells and to elucidate its characteristic. Although overexpression of hMAGEA2 in iPS cells are not different on morphology, their pluripotency and self-renewal capacity are significantly strengthened. And hMAGEA2 contributed to promote the cell cycle progression, this cell cycle changes induced proliferation acceleration. Through embryoid body formation in vitro and teratoma formation in vivo, we found that hMAGEA2 critically decreases the differentiation ability in iPS cells. Our results demonstrate that hMAGEA2 intensified the self-renewal, pluripotency, proliferation degree but efficiency of differentiaton is significantly repressed. Our findings provided that hMAGEA2 play a key role of iPS cells.

This study was conducted to investigate the effect of vitrification solution(VS) on in vitro developmental competence of immature porcine oocytes. The immature porcine oocytes were exposed to the following vitrification solution, at RT. 1) EFS sol. : 20% ethylene glycol (EG) 3 min, 40% EG + 18%(w/v) Ficoll(MV70, 000) + 0.3 M sucrose 30 sec, 2) GE sol. : 10% glycerol 5 min, 10% G + 20% EG 5 min, 25% G +25% EG 30 sec, 3) EG sol : 1.5M EG 2.5 min, 5.5 M EG + 1.0 M sucrose 30 sec. Oocytes were immediately transferred into 1.0 M, 0.5 M, 0.25 M, 0125 M, 0 M sucrose solution for 2.5 min each at RT. After removal of VS, immature oocytes were matured in vitro and subsequently all oocytes were subjected to IVF followed in vitro culture for 7 days. Maturation rates of oocytes were 38.8%, 44.5%, 22.4% and 57.6%, in EFS, EG, GE and Control, respectively, maturation rates of oocytes in EG and Control was significantly higher than EFS and GE(P<0.01). Fertilization rates of oocytes in Control was significantly higher than other treated groups(P<0.05), but no difference were observed among treated groups. Polyspermic rates were no significant difference among four groups. Cleavage rates of oocytes were 21.9%, 47.1%, 19.0% and 65.9%, in EFS, EG, GE and Control, respectively, cleavage rates of oocytes in EG and Control was significantly higher than EFS and GE(P<0.05), but blastocyst formation rates were no significant difference among four groups. These results suggested that the use of EG solution could be a great challenge for reaching a successful vitrification of immature porcine oocytes.