• Asian-Australasian Journal of Animal Sciences
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• v.11 no.6
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• pp.713-717
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• 1998

The synergistic effect of fetal calf serum (FCS) and chicken embryo extract (CEE) on protein synthesis of chicken embryo myoblasts was examined. Myoblasts were derived from chicken embryo cultured for 14 days by trypsin digestion and cultured in 5% $CO^2/95%$ air at $37^{\circ}C$. When myoblasts were cultured at the low level of cell density (20-50% of well), CEE enhanced the ability of FCS to stimulate protein synthesis of myoblasts. However, there was no significant effect of CEE to stimulate protein synthesis of myoblasts cultured at high level of cell density (100% of well).

• Reproductive and Developmental Biology
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• v.29 no.3
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• pp.145-147
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• 2005

Autocrine or paracrine mediators released by the early embryo are implicated in the support of embryonic development. Their mechanisms and optimal embryo density in the medium, however, are uncertain. This study was conducted to establish the optimal embryo density and culture medium volume in mouse parthenogenetic embryo culture. In experiment 1, culture of parthenogenetirally activated oocytes at a concentration of $2{\~}4$ embryos/${\mu}L$ significantly improved development to the blastoryst stage ($72{\%}{\leq}$) compared with culture at the lower ($0.2{\~}1$e mbryos/${\mu}L,\;0\~37.5\%$) and the higher ($5{\~}6$ embryos/${\mu}L,\;30\~53\%$) concentration for 120 h when the oocytes were cultured in a 5 ${\mu}L$ drop under mineral oil In experiment 2, the embryos cultured at a concentration of $2{\~}4$ embryos/${\mu}L$ in a 10 ${\mu}L$ drop ($81.1{\%}$) showed significantly higher blastocyst rates than those in a 5 ${\mu}L$ drop ($68.5{\%}$). This study optimizes in vitro culture condition by modifying embryo density and the volume of culture medium It may give appropriate level of autocrine and/or paracrine factors to enhance viability and subsequent normal development of mouse parthenogenetic embryos in vitro.

• Journal of Embryo Transfer
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• v.25 no.4
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• pp.259-262
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• 2010

Optimization of the preimplantation mammalian embryo culture condition was widely focused on refining medium composition under the name of chemically defined media. However, recent research revealed that the alteration of physical environment can be a crucial factor to a successful embryo development. In this study, under the same embryo density, a novel culture device named oil-free micro tube culture (MTC) system was evaluated using porcine parthenogenetic embryos. The activated oocytes were placed into the 0.2 ml thin-wall flat cap PCR tube and cultured to the blastocyst stage. As a preliminary step, embryo density and culture medium volume were optimized under a standard drop culture system. The optimal embryo density range for in vitro culture was 0.5 embryos per ${\mu}l$ in $20\;{\mu}l$ drop (20.5%) and 1.0 embryos per ${\mu}l$ in $10\;{\mu}l$ drop (20.6%). Based on these results, we compared drop culture system and 'MTC' system in terms of the developmental rate to the blastocyst stage. In $20\;{\mu}l$ medium volume, the 'MTC' system showed similar blastocyst formation rate when compared with drop culture system (20.2% versus 20.5%, respectively) while the 'MTC' system showed lower blastocyst formation rate than drop culture system in $10\;{\mu}l$ one (12.7% versus 20.0%, respectively). Therefore the $20\;{\mu}l$ MTC system may be an alternative incubation system for short-distance embryo transport without carrying the $CO_2$ incubator and this provides novel embryo culture device to clinical veterinary embryologists.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.37 no.2
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• pp.134-140
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• 1992

Carrot(Daucus carota L.) seeds (CV Danver 126) were primed and then separated by density differences to improve both the percentage and time of radicle emergence. Seeds for priming were soaked in aerated distilled water for 2 days (water imbibed), 25% solution of polyethylene glycol(PEG) 6000 for 6 or 10 days, salt solution of 0.2M KNO$_3$＋0.1M $K_2$HPO$_4$ for 6 or 10 days, or mixed with Agro-Lig with 90% moisture content for 6 days and 70% moisture content for 6 or 10 days (SMP) at 2$0^{\circ}C$, respectively. The greatest embryo growth and the highest radicle emergence were observed from the seeds treated SMP with 90% moisture content for 6 days among the primed treatments. After the SMP treatment, the seeds were separated into density classes with a float-sink procedure using aqueous solution of Maltrin 600 with 0.02/cm$^3$ density increments. The lower density classes of the carrot seeds, the more embryo growth, the higher and the faster rates of radicle emergence were exhibited in order from 1.06 to 1.14 density classes of the carrot seeds treated SMP.

• Journal of Embryo Transfer
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• v.15 no.2
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• pp.129-136
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• 2000

This study was carried out to investigate the effect of biological factors on the in vitro production(IVP) of bovine oocytes for development of simple culture methods and medium. Oocytes from the slaughterhouse ovaries were matured and fertilized using general protocol and this study was examined if there were necessary to co-culture, media change, media type and embryo density. This results were as follows: 1. The development rate according to co-culture with cumulus cells and non co-culture as drop culture was not significantly different in cleavage (68.9 vs 71.7%), 8-cell stage (41.2 vs 44.1%) and blastocyst stage (12.2 vs 13.8%), respectively (p<0.05) 2. The blastocyst development rates in YS and CRIaa were higher than that in TCM199 (12.4, 10.4$ vs 3.7%), but the cleavage (69.0, 77.8 and 61.0%) and 8-cell stage (31.7, 37.0 and 35.7%) development accoring to YS, TCM199 and CRIaa ws not significantly different, respectively (p<0.05). 3. There was no significantly different in cleavage (62.6, 59.5 and 61.2%), 8-cell(34.7, 37.9 and 34.0%) and blastocyst (9.5, 11.6 and 12.8%) development among medium change time as control, Group I and Group II, respectively (p<0.05). 4. Blastocyst formation of 8-cell stage according to embryo density was not significantly different in 1, 10 and 25 embryos (27.3, 22.5 and 34.0%), respectively (p<0.05). These results indicated that simple culture system could reduce bovine IVP embryos as drop culture as non co-culture system, high density embryo (25 embryos/50 $\mu$1 drop). YS defined medium and no medium change for whole culture period, although other biological factors need to examine in order to produce efficient IVP bovine embryos.

• Journal of Embryo Transfer
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• v.15 no.3
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• pp.271-277
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• 2000

This study was conducted to establish the optimal culture conditions for in vitro production of bovine embryos derived from slaughter house ovaries. Cumulus-oocyte- complexes (COCs) collected by aspiration from follicles of 2~7 mm in diameter were matured in Ham's F-10 medium supplemented with 0.01 $\mu\textrm{g}$/m1 epidermal growth factor (EGF) at 39$^{\circ}C$ in a humidified atmosphere of 5% $CO_2$in air. After 24 hrs of culture, the oocytes were co-cultured with epididymal sperm selected off by Percoll-density gradient in TALP medium for 24 hrs. The presumptive zygotes were cultured in HECM-6 medium for 3 d post-insemination, and followed by cultured in TCM199 medium until 7 to 10d post-insemination. The cultures were compared of their cleavage and development into later stage in culture medium by additions of different protein sources (PVA, BSA and BCS) and by different embryo density. The rates of cleavage and development rates into blastocyst were not significantly (P<0.05) different among the culture media containing with BSA (75.0% and 40.5%), BCS (76.7% and 38.0%) and PVA (72.5% and 42.2%), respectively. Significantly (P<0.05) higher blastocysts rates were obtained in culturing of 30 and 40 embryos in each 50$\mu$l droplets of culture medium than in 5, 10 and 20 embryos. These results indicate that the optimal density of embryos is 30~40 embryos in a 50$\mu$l droplet of culture medium. Furthermore there is no effect of different protein sources on early embryonic development.

• Applied Biological Chemistry
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• v.51 no.3
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• pp.200-206
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• 2008

We investigated the effect of rice embryo and embryo jelly with black rice bran pigment on lipid metabolism and antioxidant activity. Thirty 4-week-old male Sprague-Dawley rats were fed high cholesterol diets supplemented with 15% rice embryo and 25% embryo jelly added black rice bran pigment, respectively, for 6 weeks. Plasma and hepatic lipid profile, lipid peroxidation, and the activity of antioxidant scavenger enzymes in liver were examined. Supplementation with rice embryo and embryo jelly had no effect on food intakes in high cholesterol-fed rats. The plasma triglyceride concentration was not significantly different among the groups. Supplementation with rice embryo and embryo jelly resulted in lower plasma and hepatic total cholesterol (TC) concentration and high-density lipoprotein-cholesterol (HDL-C)/TC ratio and atherogenic index compared to the control group, while the plasma HDL-C concentration tended to elevated. Rice embryo and embryo jelly tended to lower plasma and hepatic levels of thiobarbituric acid reactive substances than the control group. Moreover, hepatic antioxidant enzyme activities, including superoxide dismutase and glutathione peroxidase, were significantly higher in the rice embryo and embryo jelly groups. In conclusion, rice embryo and embryo jelly was very effective in improving the lipid metabolism and reducing oxidative stress by up-regulating the hepatic antioxidant enzymes in high cholesterol-fed rats.

• Journal of Plant Biotechnology
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• v.49 no.1
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• pp.61-73
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• 2022

In this study, we developed a reliable and efficient Agrobacterium-mediated genetic transformation system by applying sonication and vacuum infiltration to six chickpea cultivars (ICCV2, ICCV10, ICCV92944, ICCV37, JAKI9218, and JG11) using embryo axis explants. Wounded explants were precultured for 3 days in shoot induction medium (SIM) before sonication and vacuum infiltration with an Agrobacterium suspension and co-cultivated for 3 days in co-cultivation medium containing 100 µM/l of acetosyringone and 200 mg/l of L-cysteine. Responsive explants with putatively transformed shoots were selected using a gradual increase in kanamycin from 25 mg/l to 100 mg/l in selection medium to eliminate escapes. Results showed optimal transformation efficiency at a bacterial density of 1.0, an optical density at 600 nm wavelength (OD₆₀₀), and an infection duration of 30 min. The presence and stable integration of the β-glucuronidase (gusA) gene into the chickpea genome were confirmed using GUS histochemical assay and polymerase chain reaction. A high transformation efficiency was achieved among the different factors tested using embryo axis explants of cv. JAKI 9218. Of the six chickpea cultivars tested, JAKI9218 showed the highest transformation efficiency of 8.6%, followed by JG11 (7.2%), ICCV92944 (6.8%), ICCV37 (5.4%), ICCV2 (4.8%), and ICCV10 (4.6%). These findings showed that the Agrobacterium-mediated genetic transformation system will help transfer novel candidate genes into chickpea.

• Journal of Plant Biology
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• v.35 no.1
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• pp.45-51
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• 1992

The changes of protein bodies in endosperm cells of both seeds with red seed coat and indehiscent seeds of Panax ginseng C.A. Meyer have been investigated in relation to the embryo development. In the early stage of seeds with red seed coat, spherical spherosomes were distributed in endosperm cells. Protein bodies were formed from vacuoles containing the storage protein. Cell organelles were hardly observed in the cytoplasm. In the late stage of the seed with red seed coat, the endosperm was filled with spherosomes and protein bodies. The protein bodies consisted of amorphous inclusions with high electron density or proteinaceous matrix with even electron density. In the seed of in dehiscence, the protein body in endosperm cells contained globoids and protein crystalloids. The globoid of protein body had a electron dense materials. Umbiliform layer was formed between embryo and endosperm. The deformation patterns of endosperm cell wall and the cellulose microfibril were observed in endosperm cells near the umbiliform layer. Umbiliform layer consisted of lipid body and autolyzed cell debris. The protein body of endosperm cell near the umbiliform layer showed various degenerative patterns, and so electron density of proteinaceous matrix was gradually decreased.reased.

• Environmental Analysis Health and Toxicology
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• v.22 no.3
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• pp.211-218
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• 2007

Even though some standard developmental bioassay protocols for environmental assessment using sea urchins have already been described, there have not been many attempts to apply and modify these protocols with Korean species. Therefore, there is a strong need to establish standard bioassay protocols using sea urchins commonly found in Korea. Prior to developing a new protocol, it is essential to know the optimal conditions for the bioassay procedures. We investigated the optimal conditions (temperature, salinity, and embryo density) of the sea urchin Strongylocentrotus intermedius. The ideal temperature for developmental bioassay of S. intermedius was determined to be $15^{\circ}C$ and the time required for the embryo to become pluteus larva was 72 hr. The optimal range of salinity for the embryo toxicity using S. intermedius was between 30 to 32 psu, which is similar to the range found in the natural habitats of adult populations. The optimum density of embryos at the beginning of bioassays was 100 embryos/mL. When the assays were carried out at higher densities, the proportion of normally developed larvae decreased significantly.