• Development and Reproduction
• /
• v.8 no.2
• /
• pp.119-122
• /
• 2004

It is well known that unidentified factors in sera, hormones and growth factors promote the proliferation of granulosa cells and nuclear maturation of bovine COCs in vitro. Attemps had been developed the simple composition of culture media and similar system to in vivo conditions has been applied. In the present study, we investigated the effect of TGF-${\beta}$ on in vitro maturation and in vitro development of Hanwoo COCs. When the COCs were matured in TCM 199 containing 0.1, 1 or 10 ng/ml TGF-${\beta}$ for 24 hrs, metaphaseⅡ of COCs were obtained 95.8%, 100% of matured COCs, respectively and there were no differences among the concentrations of TGF-${\beta}$. Matured COCs with TGF-${\beta}$ cultured in maturation medium after in vitro fertilization, developmental rate to blastocyst were 0~0.8%. Matured COCs with TGF-${\beta}$ were cultured in TCM 199+10% FBS, 0.8% BSA, 0.1% PVA, blastocyst formation were showed in 12.4%, 12.8%, 8.5% of those and cultured in IVMD or IVD without serum were 38.4%, 34.8%, respectively. There were significant differences among the media (P<0.05). TGF-${\beta}$ is available for i vitro maturation of bovine COCs, but further investigation would be need for finding the synergistic autocrine/paracrine fashion of other growth factors in early bovine development.

• Journal of Bio-Environment Control
• /
• v.28 no.2
• /
• pp.110-116
• /
• 2019

This study was conducted to determine the effects of a pre-planting fertilizers with various $NH_4{^+}:NO_3{^-}$ ratios in a coir dust:peatmoss:perlite (3.5:3.5:3.0, v/v/v) medium on the growth of hot pepper (Capsicum annuum L. cv. Nokkwang) plug seedling. Nitrogen levels were fixed to $300mg{\cdot}L^{-1}$ and the $NH_4{^+}:NO_3{^-}$ ratios were varied to 0:100, 27:73, 50:50, 73:27, and 100:0. The 50-cell trays were filled with treatment media containing pre-plant fertilizers, then seeds were sown. After seeds were germinated, the trays were moved to greenhouse and seedlings were feed with 13-2-13 and 20-9-20 fertilizers, alternatively. The changes in pH and EC were measured every week and soil solution for nutrient concentrations were analyzed in week 0, 3, and 7. The measurements of seedling growths as well as analysis of tissue nutrient contents were also conducted in week 7. The varied $NH_4{^+}:NO_3{^-}$ ratios did not influence on the pHs of root media after incorporation of pre-planting fertilizers, but the ECs were heightened as proportion of $NH_4{^+}$ to $NO_3{^-}$ were elevated. During the raising of seedlings, the pHs rose over time in the treatments of 0:100 and 27:73 ($NH_4{^+}:NO_3{^-}$). The concentrations of all macro-elements in root media decreased gradually as seedlings grew in all treatments. The seedling growths 7 weeks after seed sowing were the highest in the treatments of 27:73 and 50:50 ($NH_4{^+}:NO_3{^-}$) and those became worse in the treatments of higher $NH_4{^+}$ ratios than 73%. In terms of inorganic element contents based on the dry weight of above ground tissue, the treatment of 0:100 showed the lowest content of Ca, Mg, Na, Cu, Mn, and Zn. Based on the results, it is desired that $NH_4{^+}$ ratio in pre-planting fertilization is maintained to be 50% or less for the raising of hot pepper plug seedlings.

• Korean Journal of Animal Reproduction
• /
• v.22 no.4
• /
• pp.425-435
• /
• 1998

To improve the efficiency of nuclear transplantation in bovine, in this study the development in vitro of nuclear transferred (NT) embryos was compared by different activation regimens of the enucleated oocytes. The effect of developmental stage and culture system of donor nuclei on fusion and development in vitro of NT embryos were also evaluated. Oocytes were collected from Hanwoo ovaries obtained from slaughterhouse and matured in Ham's F-10 supplemented with hormones. After 20~22 h maturation, the oocytes were vortexed to be free from cumulus cells and subsequently their nucleus and the first polar body were removed. Enucleated oocytes were divided into 3 groups for activation; the oocytes of group I were activated with ionomycin for 5 min and subsequently incubated in 6-dimetylarninopurine (DMAP) for 4 h, Those of group II were treated with DMAP for 4 h at 39 h after onset of in vitro maturation (IVM) and those of group III were kept in room temperature ($25^{\circ}C$) for 3 h at 39 h after onset of IVM. After in vitro fertilization (IVF) the embryos for muclear donor were cultured either by group culture (20 embryos /50 ${mu}ell$ drop) or individually (1 embryo /50 ${mu}ell$ drop) for 4 day and 5 day. At day 4 and 5 after IVF, blastomeres were separated in calcium-magnesium free medium, and then classified into small (day 5: $\leq$ 38 ${\mu}{\textrm}{m}$, day 4: $\leq$ 46 ${\mu}{\textrm}{m}$) and large (day 5 : $\geq$ 38 ${\mu}{\textrm}{m}$, day 4 ; $\geq$ 46 ${\mu}{\textrm}{m}$). The separated blastomeres were replaced into enucleated and activated recipient cytoplasm. The blastomere-oocyte complexes were fused by electrically. The NT embryos were cultured in TCM-199 containing 10% FCS in 39$^{\circ}C$, 5% $CO_2$ incubator for 7 day. The results obtained were summarized as follows; There were no differences in fusion and development to blastocyst between groups as group I (68%, 10%), group II (75%, 14%) and group III (73%, 9%), respectively. However, the cell number in blastocyst of NT embryos in group III were significantly fewer than in the other groups (P＜0.05). No differences in fusion and development to blastocyst were found between individual or group cultured and between small or large blastomeres of day 4 and day 5 donor embryos. From these results, it was concluded that the combination of ionomycin and DMAP, or treatment of DMAP at 39 h after onset of IVM were useful for the efficient of production of NT bovine embryos, and the individual cultured embryos could be simply used as donor nuclei for NT bovine embryo.