| dc.description.abstract | This study was conducted with two objectives: First, establish in vitro
maturation culture system of porcine oocyte and then examine effect of
caffeine treatment on aging oocytes. Although in vitro maturation culture
system was established in some advanced technology Laboratories in the
world, it has not been established well in Vietnam. Using experience
working in porcine oocyte, I established a culture system with 90% of
oocyte acquire meiotic competence (Metaphase I: 12.7%, Anaphase ITelophase I: 10.3 % and Metaphase II: 65.3%, Table 1). The culture
medium using in this study was bicarbonate-buffered TCM-199
supplemented with 10% (v/v) fetal bovine serum (FBS) (GIBCO, life
technologies, US), 0.1 mg/ml sodium pyruvate (Sigma-Aldrich Co, LLC,
Japan), 10% (v/v) porcine follicular fluid (pFF), 0.1 IU/ml human chorionic
gonadotropin (hCG) (Sigma, USA). I found that the important role of 10%
porcine follicular fluid increased number of mature oocyte (MII) with high
quality. These mature oocytes showed histone H3 deacetylation as
previous reports. Then, I examined the expression of histone H3
acetylation in aging oocyte by extended culture of mature oocyte for 24h.
Here, I demonstrated that the aging porcine oocytes have re-acetylation of
histone H3 at lysine 14 (Fig. 4), similar phenomenal was reported in
mouse. Finally, to extend my study by using caffeine to prevent aging
oocyte, MII porcine oocytes were treated with 2.5 mM caffeine for 45 min
and incubate in TCM-199 for 24h. The treated oocyte expressed reacetylation of histone H3 at lysine 14. We suggested that the present
concentration and duration of treated caffeine were not enough for
preventing aging of porcine oocytes.
Keywords: in vitro maturation, pig oocyte, histone acetylation, aging, caffeine | en_US |
