Biomedical subjects

Il-Keun Kong

Publications and source records attributed to Il-Keun Kong.

6 recordsLinked to original sources

PubMed2026

Epigenetic reduction OF H3K9me3 and H3K27me3 by RK-701 and GSK 126 improves the developmental competence of bovine SCNT embryos.

Somatic cell nuclear transfer (SCNT) failure has largely been attributed to incomplete epigenetic reprogramming, particularly the dysregulation of repressive histone modifications such as H3K9me3 and H3K27me3. Reducing these repressive marks has been shown to improve reprogramming efficiency in SCNT embryos. Although histone demethylase mRNA injection has been used for this purpose, it is labor-intensive, technically demanding, and time-consuming. In this study, we investigated a simplified approach that combined RK-701 and GSK-126 to reduce H3K9me3 and H3K27me3 levels, respectively, in bovine SCNT embryos. Three experimental groups were established: IVF embryos (control), SCNT-control (SCNT-C) embryos, and inhibitor-treated SCNT embryos (SCNT-T). The IVF group was used as a reference standard. Fused one-cell SCNT embryos were treated with 2 μM RK-701 and 0.2 μM GSK-126 from the one-cell stage to the 16-cell stage. Gene expression analysis at the 16-cell stage revealed a significant reduction in histone methyltransferase (HMT) expression (p < 0.05), and immunofluorescence analysis confirmed marked decreases in H3K9me3 and H3K27me3 levels. In addition, the expression of genes associated with zygotic genome activation (ZGA) and pluripotency was significantly higher in SCNT-T embryos than in SCNT-C embryos. Assessment of blastocyst quality revealed reduced reactive oxygen species (ROS) levels, decreased expression of apoptosis-related genes, and improved mitochondrial membrane potential in the treated group, as indicated by JC1 staining. Overall, this approach effectively reduced repressive histone marks, enhanced epigenetic reprogramming, and improved ZGA, thereby increasing the developmental rate and adhesion potential of bovine SCNT embryos. These findings suggest that combined treatment with RK-701 and GSK-126 may provide a simple and practical strategy for improving the efficiency of bovine cloning.

Bovine embryos↗

PubMed2006

Effect of serum starvation on the efficiency of nuclear transfer using odd-eyed white cat fibroblasts.

In the present study, we compared in vitro and in vivo development of nuclear transfer (NT) embryos derived from serum-starved or non-serum-starved odd-eyed cat skin fibroblast cells. Flow cytometry analyses revealed that a higher percentage of cells were in the G0/G1 phase after serum starvation (89.3%) as compared with non-serum-starved cells (73.8%, P<0.05). Frequency of cleavage and development to the blastocyst stage was not different between the serum-starved or non-serum-starved treatment group, 67.9 and 12.5% versus 73.0 and 10.2%, respectively (P>0.05). After transfer of two to four-cell NT embryos derived from starved and non-starved fibroblasts, three of nine (33%) and one of nine (11%) recipients delivered three live male (plus, one dead) and two live male kittens, respectively. Of the five live-cloned kittens, one died from diarrhea at 3 weeks of age and the other four kittens are growing at a normal rate. The cloned kittens are blue-eyed and have functional auditory systems, including clones of the odd-eyed deaf Turkish Angora cat. Subsequent DNA analysis of nine-cat specific microsatellite loci confirmed that all of the cloned kittens were identical to the odd-eyed donor male, but a point mutation occurred in the dead fetus at the FCA 290 marker.

Animals↗

PubMed2006

Anticoccidial effect of green tea-based diets against Eimeria maxima.

Anticoccidial effects of green tea (GT)-based diets were evaluated in chickens following oral infection with Eimeria maxima an ubiquitous intestinal parasite of poultry that impairs the growth and feed efficiency of infected birds. Five-week-old chickens were assigned to four groups (GT 0.5%, GT 2.0%, untreated/infected and non-infected control) and each group consisted of 15 chickens. Chickens were fed a standard diet supplemented with ground green tea for 2 weeks prior to infection with E. maxima (10,000 sporulated oocysts per bird). The effects of green tea on E. maxima infection were assessed by two parameters, fecal oocyst shedding and body weight gain. The green tea-fed chickens produced significantly reduced fecal oocysts (P<0.05) when compared to the E. maxima-infected group fed standard diet. The green tea-based diet, however, did not improve body weight loss caused by E. maxima infection. This study is the first to demonstrate anticoccidial effect of green tea on Eimeria parasites.

Animal Feed↗

PubMed2006

Epidermal growth factor enhances meiotic resumption of canine oocytes in the presence of BSA.

Despite many attempts to improve the in vitro maturation (IVM) of canine oocytes using various culture conditions, the efficiency of canine IVM remains very low compared with that of other domestic animals. In the present study we examined the effect of ovarian estrus stage on oocyte quality, and the effect of epidermal growth factor (EGF) in the presence and absence of macromolecules on the IVM of canine oocytes. More oocytes >or=100 microm in diameter were obtained from follicular ovaries than from ovaries at other estrus stages. After 72 h of culture, significantly more oocytes recovered from follicular ovaries than from anestrous and luteal ovaries were in germinal vesicle break down (GVBD). Bovine serum albumin (BSA) or fetal bovine serum (FBS) supplementation improved meiotic resumption as compared to polyvinyl alcohol (PVA) supplementation; however, there was no difference between the BSA and FBS supplements. The oocytes matured in North Carolina State University (NCSU) 37 medium containing 0.4% BSA and 100 ng/ml EGF showed the highest rates of development to the metaphase II (MII) stage when compared with the control treatment (P < 0.05). These results suggest that the estrous cycle of bitches influences the meiotic resumption of oocytes cultured in vitro, and EGF increases the meiotic resumption of canine oocytes in the presence of BSA in vitro.

Animals↗

PubMed2006

Chromatin, microtubule and microfilament configurations in the canine oocyte.

In the present study, we observed chromatin, microtubule and microfilament distribution in canine oocytes. The germinal vesicle (GV) chromatin of canine oocytes was classified into four configurations (GV-I, -II, -III and -IV) based on the degree of chromatin separation and condensation. Oocytes recovered from follicular phase ovaries had a greater amount (68%, P < 0.05) of GV-III or GV-IV chromatin than did those from non-follicular phase ovaries (35%). The majority (86.7%) of in vivo ovulated oocytes were at GV-IV. The rates of development to GV breakdown/metaphase I/metaphase II were higher in oocytes recovered from follicular ovaries than from non-follicular ovaries. Immunostaining results revealed cytoplasmic microtubules present in all GV-stage oocytes. Following GV breakdown, microtubular asters were produced from condensed chromatin. The asters appeared to be elongated, and encompassed condensed chromatin particles to form meiotic metaphase chromatin. Microfilaments were located in the cortex and around the GV. During meiotic maturation, a microfilament-rich area, in which the chromatin is allocated, was observed in the oocyte. Our results indicate that oocytes recovered from follicular ovaries were in an advanced stage of GV, and were more competent to complete maturation compared to those from non-follicular phase ovaries. Both microtubules and microfilaments are closely associated with reconstruction of chromatin during meiotic maturation in canine oocytes.

Actin Cytoskeleton↗

PubMed2002

Improvement in post-thaw viability of in vitro-produced bovine blastocysts vitrified by glass micropipette (GMP).

The purpose of this study was to investigate the use of a glass micropipette (GMP) as a vessel for vitrification of in vitro-produced (IVP) bovine blastocysts and to compare the results with post-thaw survival rate of bovine blastocysts frozen in GMP with those frozen in open pulled straw (OPS) that have been previously investigated. The GMP vessel permitted higher freezing and warming rates than the OPS due to the higher heat conductivity of the glass and the lower mass of the solution that contained the embryos. Groups of three bovine IVP blastocysts were sequentially placed into vitrification solution before being loaded into either the OPS or GMP vessels and they immersed into LN(2) within 20-25 s. Post-thaw blastocysts were serially washed in 0.25 and 0.15 M sucrose in a holding medium (HM: D-PBS supplemented with 5% FCS) and then in TCM-199 for 5 min in both cases. They was then cultured in TCM 199 supplemented with 10% FCS for 24 or 48 h. The rate of blastocyst re-expansion was significantly different between OPS (79.6%) and GMP (90.4%) methods. Neither was the hatching rate significantly different among OPS (51.8%), GMP (57.1%) methods and non-vitrified group (67.3%). Only the rate of post-thaw re-expanding of blastocysts loaded in narrow column was significantly higher than that of the wide column (83.3% versus 56.7%) (P < 0.05), although the GMP straw was loaded with three blastocysts per vessel. These results indicated that the GMP vessels provided high survival rates of bovine IVP blastocysts. The location of the embryos loaded into a narrow or wide portion was considered to be a limiting factor to the viability of bovine IVP embryos.

Animals↗