• Title/Summary/Keyword: in vitro effect

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Successful 20 hours Canine Allograft Preservation with new Solution Containing Triiodothyronine - Development of new lung preservation solution II - (삼요드티로닌을 포함한 폐보존액을 이용한 20시간 폐보존 - 새로운 폐 보존액의 개발 II -)

  • 성숙환;김영태;김주현
    • Journal of Chest Surgery
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    • v.32 no.5
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    • pp.413-421
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    • 1999
  • Background: Ischemia reperfusion injury is known to contribute to the major causes of the early graft failure in lung transplantation. Triiodothyronine (T3) has been suggested to ameliorate ischemia reperfusion injury from both in vivo and in vitro experiments of various organs. Prospecting its beneficial effect for pulmonary allograft preservation, we made a new solution by adding T3 into the extracellular type dextran solution. Material and Method: Twelve adult mongrel dogs underwent left lung allotransplantation. Six donor dogs were flushed with the new solution(Group 1, n=6), and the remaining six were flushed with Euro-Collins solution to serve as controls(Group 2, n=6). Allografts were stored in each preservation solution for 20 hours at 4$^{\circ}C$. Left single lung transplantations were performed. The right pulmonary artery and the right main bronchus were clamped at 15 minutes after the reperfusion and maintained throughout the experiment to evaluate the transplanted left lung function. Result: Arterial carbon dioxide tension was better in group 1 than in group 2 throughout the experiment period and the difference was statistically significant at 2 hours after reperfusion(28.0${\pm}$3.0 mmHg and 53.1${\pm}$17.4 mmHg, p<0.05). The differences of arterial oxygen partial pressure, peak airway pressure and pulmonary vascular resistance showed no statistical significance. The malondialdehyde(MDA) level, measured from tissue obtained at 120 minutes after reperfusion showed no statistically significant difference. The tissue wet/dry ratio of group 1(649${\pm}$27 %) was significantly lower than that of group 2(686${\pm}$71 %, p<0.05). The microscopic examination revealed varying degrees of injury represented mainly by findings such as perivascular neutrophil infiltration, capillary hemorrhage and interstitial congestion. These findings were less severe in group 1 than those in group 2. Conclusion: The new solution demonstrated superior allograft preservation after 20 hour ischemia compared to Euro-Collins solution in canine single left lung transplantation model, these results suggest that T3 might be a promising agent for pulmonary allograft preservation.

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Protective Effects of Perilla frutescens Britt var. japonica Extracts from Oxidative Stress in Human HaCaT Keratinocytes (HaCaT 피부각질세포에서 들깻잎 추출물의 산화적 스트레스에 대한 항산화 효과)

  • Ji, Na;Song, Jia-Le;Kil, Jeung-Ha;Park, Kun-Young
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.42 no.2
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    • pp.161-167
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    • 2013
  • The aim of this study was to investigate the protective effects of methanolic extract from perilla (Perilla frutescens Britt var. japonica) leaves (PLME) on oxidative injury from hydrogen peroxide ($H_2O_2$) in human HaCaT keratinoctyes. Cells were co-incubated with various concentrations (0~200 ${\mu}g/mL$) of PLME for 24 hr, and then exposed to $H_2O_2$ (500 ${\mu}M$) for 4 hr. $H_2O_2$ significantly decreased cell viability (p<0.05). However, PLME provided protection from $H_2O_2$-induced HaCaT cell oxidation in a dose-dependent manner. To further investigate the protective effects of PLME on $H_2O_2$-induced oxidative stress in HaCaT cells, the cellular levels of lipid peroxidation, and antioxidant enzymes (including superoxide dismutase (SOD), glutathione peroxidase (GSH-px) and catalase (CAT)) were measured. PLME decreased cellular levels of lipid peroxidation, and also increased the activities of antioxidant enzymes. In addition, the antioxidant activities of PLME were also determined by DPPH and hydroxyl (${\cdot}OH$) radical scavenging assay, and major antioxidant compounds of PLME were measured by colorimetric methods. DPPH and ${\cdot}OH$ radical scavenging activities of PLME increased in a dose dependent manner and was similar to the DPPH scavenging activity of ascorbic acid at 50 ${\mu}g/mL$; however PLME activities were stronger than ascorbic acid (50 ${\mu}g/mL$) in the ${\cdot}OH$ scavenging assay. The amounts of antioxidant compounds, including total polyphenolics, total flavonoids, and total ascorbic acid from PLME were $52.2{\pm}1.1$ mg gallic acid (GAE)/g, $33.7{\pm}4.7$ mg rutin (RUE)/g, and $17.0{\pm}0.5$ mg ascorbic acid (AA)/g, respectively. These results suggest that PLME has a strong free radical-scavenging activity and a protective effect against $H_2O_2$-induced oxidative stress in the keratinocytes.

Bifidogenic Effects of Inuloprebiotics in Broiler Chickens (이눌로프리바이오틱스의 브로일러에 대한 비피더스균 활성 효과)

  • Park, Byung-Sung
    • Journal of Life Science
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    • v.18 no.12
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    • pp.1693-1699
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    • 2008
  • Recent studies have suggested that inulin might be utilized as a prebiotics for the promotion of antimicrobial growth, but a major obstacle to the use of inulin has been its low bifidogenic effects, which were initially observed in the ceca of broiler chickens. Inulin has some problems with related to denaturation in air and lowering passage rate from upper digestive tract to caecum. To solve this problems, a newly developed compound derived by microencapsulation, inuloprebiotics, was hypothesized to enrich cecal bifidobacterial populations and reduce the colonization levels of Salmonella in the ceca of broiler chickens. The in vitro growth of intestinal beneficial bacteria including Bifidobacterium longum, Bifidobacterium bifidum, Lactobacillus acidophilus, and Lactobacillus casei grew effectively on the medium containing inulin, whereas the growth of Streptococcus aureus and Clostridium perfringens was not differences among the treatment groups. Broiler chickens consumed chow diets containing 0.5%, 0.7% or 1.0% inuloprebiotics, or a control diet without inuloprebiotics supplementation. The chickens on the inuloprebioticssupplemented diets evidenced significantly higher cecal levels of Bifidobacterium and Lactobacillus species as compared with the chickens on the control diet. The population of cecal E. coli and Salmonella was specifically reduced as the result of treatment with inuloprebiotics. However, we noted no significant differences in Bifidobacterium species, E. coli and Salmonella counts among the inuloprebiotics treatment groups. The inuloprebiotics-supplemented diets induced an increase in the serum IgG concentration. The thymus index was significantly increased in the broiler chickens that consumed diets containing 0.7% or 1.0% inuloprebiotics, with the exception of the chickens consuming the diet supplemented with 0.5% inuloprebiotics. These results indicate that the inuloprebiotic preparations exerted an immune system-promoting effect or selectively enriched the cecal Bifidobacterium species populations in the broiler chickens, and also suggest that inuloprebiotics may prove useful as a stable natural antimicrobial agent.

Lipase-Inhibitory and Anti-Oxidative Activity of the Methanol Extract and the Powder of Phellinus linteus (상황버섯 자실체 메탄올 추출물과 분말의 지방소화효소 억제 및 항산화 활성)

  • Kim, Ji-Hyun;Son, In-Suk;Kim, Jong-Sang;Kim, Ki-Hoon;Kwon, Chong-Suk
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.37 no.2
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    • pp.154-161
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    • 2008
  • Phellinus linteus (PL) has been known to exhibit potent biological activity. The present study was designed to investigate lipase-inhibitory and anti-oxidative activity of the methanol extract and the powder of PL fruiting body. The methanol extract of PL appeared to have the inhibitory activity against pancreatic lipase with an $IC_{50}$ value of $36.3\;{\mu}g/mL$, and the scavenging activity of DPPH radical with an $IC_{50}$ value of $20.1\;{\mu}g/mL$, which was similar to that of vitamin C ($IC_{50}\;18.3\;{\mu}g/mL$). To investigate the lipase-inhibitory and anti-oxidative effect of PL on animal, Sprague-Dawley rats were fed with high-fat diet supplemented with either 2% or 5% PL powder for 8 weeks. Total food intake was significantly increased, but body weight was not changed by PL powder supplementation. However, fecal fat excretion of the experimental groups fed with the PL powder were higher than that of the control group. PL powder showed a decrease in the plasma total cholesterol, LDL-cholesterol, and the hepatic total cholesterol levels. The anti-oxidative enzyme activities were also affected by PL supplementation. Glutathione peroxidase (GSH-Px) in the plasma and liver were significantly increased by 98% and 46% in the 2% PL group, and 99% and 32% in the 5% PL group, respectively. Total superoxide dismutase (T-SOD) activity was not affected by PL supplementation. DNA damage was measured by the comet assay in the lymphocytes collected after 2 weeks, 4 weeks, and 8 weeks of feeding PL supplemented diet. Lymphocyte DNA damage was decreased in the PL supplemented group. Furthermore, PL feeding enhanced the resistance to lymphocyte DNA damage caused by an oxidant challenge with $H_2O_2$.

Growth enhancement and cytotoxicity of Korean mistletoe fractions on human cell lines (한국산 겨우살이 분획물의 면역세포의 생육증진 및 세포독성)

  • Lee, So-Jin;Lee, Mi-Kyoung;Choi, Geun-Pyo;Yu, Chang-Yeon;Roh, Seong-Kyu;Kim, Jong-Dai;Lee, Hyeon-Yong;Lee, Jin-Ha
    • Korean Journal of Medicinal Crop Science
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    • v.11 no.1
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    • pp.62-70
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    • 2003
  • The biological activities on human immune and cancer cell lines of the four kinds of Korean mistletoes (Korean Viscum album, var. coloratum, : Korean Viscum sp. in Quercus acutissima Carr., Korean Viscum sp. in Castanea crenata, Korean Viscum sp. in Betula platyphylla, and Korean Viscum sp. in Salix koreensis) extracts were investigated. The extracts were preparated with ethanol, and fractionated with n-butanol, ethyl acetate, chloroform, hexane, and second distilled water. Cytotoxic potencies of the fractions on human normal lung cell line (HEL 299) showed under 28% in the concentration of 0.5 mg/ml. Growth inhibition effect of the Korean mistletoe extracts on the several human cancer cell lines depends on the concentration of the extracts, and extracting solvent. The hexane, chloroform, and ethyl acetate fractions indicated a strong anticancer activity, but not in aqueous and butanol fractions. Some mistletoe fractions have a different characteristic on the cancer cell lines. Stimulation on the growth of human immuno cell lines(B cell : Raji, T cell: Jurkat) of the extracts were confirmed in the ethyl acetate, chloroform, hexane fractions, but not in aqueous system.

A Study on the Antioxidative, Anti-inflammatory and Anti-thrombogenic Effects of Zanthoxylum piperitum DC. Extract (초피나무 추출물의 항산화, 항염증 및 항혈전 효능에 관한 연구)

  • Jang, Mi-Jin;Rhee, Soon-Jae;Cho, Sung-Hee;Woo, Mi-Hee;Choi, Jeong-Hwa
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.35 no.1
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    • pp.21-27
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    • 2006
  • Effects of root, stem and leaf extracts of Zanthoxylum piperitum on the inhibition of lipid peroxidation in the hepatic microsome of rat, DPPH radical scavenging activity, soybean lipoxygenase activity and activated partial thromboplastin times (APTT) were examined in vitro. The highest inhibition of hepatic microsomal lipid peroxidation was observed by ethyl acetate fraction of the root and stem extracts. The high inhibition of lipid peroxidation was observed in the leaf, the root and the stem in order. The DPPH radical scavenging activity of ethyl acetate fraction was higher than that of n-butanol fraction and it was similar to the root and the steam extract. It was similar to the inhibition of hepatic microsomal lipid peroxidation. The DPPH radical scavenging activity was the highest in 0.50 mg/mL of ethyl acetate fraction, and it was 4.4-fold higher than that of a-tocopherol, as an antioxidant standard. The DPPH radical scavenging activity was dependent on the extract concentration in the range of $0.12\~5.00$ mg/mL. The soybean lipoxygenase activity of ethyl acetate fraction was higher than that of n-butanol fraction and it was similar to the root and the stem extracts. The soybean lipoxygenase activity was the highest in 0.50 mg/mL of ethyl acetate fraction. The soybean lipoxygenase activity was dependent on the extract concentration in the range of $0.12\~5.00$ mg/mL. The leaf extract showed the highest antithrombogenic effect followed by the stem and then the root extract. The activated partial thromboplastin times were dependent on the extract concentration in the range of $0.10\~2.00$ mg/mL.

The effect of tumor necrosis factor (TNF)-α to induce matrix metalloproteinase (MMPs) from the human dental pulp, gingival, and periodontal ligament cells (사람의 치수, 치은, 치주인대 세포에 tumor necrosis factor (TNF)-α로 자극 시 matrix metalloproteinase (MMPs)의 분비에 관한 연구)

  • Rhim, Eun-Mi;Park, Sang-Hyuk;Kim, Duck-Su;Kim, Sun-Young;Choi, Kyoung-Kyu;Choi, Gi-Woon
    • Restorative Dentistry and Endodontics
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    • v.36 no.1
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    • pp.26-36
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    • 2011
  • Objectives: In the present study, three kinds of tissues cells (pulp, gingiva, and periodontal ligament) were investigated if those cells express MMP and TIMP when they were stimulated with neuropeptides (substance P, CGRP) or proinflammatory cytokine, TNF-$\alpha$. Materials and Methods: The cells cultured from human dental pulp (PF), gingiva (GF) and periodontal ligament were (PDLF) stimulated with Mock, SP, TNF-$\alpha$, and CGRP for 24 hrs and 48 hrs. for an RNase protection assay and Enzyme Linked Immunosorbent Assay. Cells (PF, GF and PDLF) seeded in 100 mm culture dish were stimulated with SP ($10^{-5}$, $10^{-8}\;M$) or only with medium (Mock stimulation) for 4hrs and for 24 hrs for RNase Protection Assay, and they were stimulated with CGRP ($10^{-5}\;M$) and TNF-$\alpha$(2 ng/mL) for 24 hrs and with various concentraion of TNF-$\alpha$(2, 10, and 100 ng/mL) for Rnase Protection Assay with a human MMP-1 probe set including MMP 1, 2, 8, 7, 8, 9, 12, and TIMP 2, 3. In addition, cells (PF, GF and PDLF) were stimulated with Mock and various concentraion of TNF-$\alpha$(2, 10, and 100 ng/mL) for 24 hrs and with TNF-$\alpha$(10 ng/mL) for 48 hrs, and the supernatents from the cells were collected for Enzyme Linked Immunosorbent Assay (ELISA) for MMP-1 and MMP-13. Results: The expression of MMPs in PF, GF, PDLF after stimulation with SP and CGRP were not changed compared with Mock stimulation for 4 hrs and 24 hrs. The expression of MMP-1, -12, -13 24 hrs after stimulation with TNF-$\alpha$ were upregulated, however the expression of TIMP-3 in PF, GF, PDLF after stimulation with TNF-$\alpha$ were downregulated. TNF-$\alpha$(2 ng/mL, 10 ng/mL, 100 ng/mL) increased MMP-1 and MMP-12 expression in PF dose dependently for 24 hrs. Conclusions: TNF-$\alpha$ in the area of inflammation may play an important role in regulating the remodeling of dentin, cementum, and alveolar bone.

Hepatoprotective Effects of Semisulcospira libertine Hydrolysate on Alcohol-induced Fatty Liver in Mice (알코올성 지방간 유발 마우스에서 다슬기 유래 가수분해물의 간 보호 효과)

  • Song, Eun Jin;Cho, Kyoung Hwan;Choo, Ho Jin;Yang, Eun Young;Jung, Yoon Kyoung;Seo, Min Gyun;Kim, Jong Cheol;Kang, Eun Ju;Ryu, Gi Hyung;Park, Beom Yong;Hah, Young-Sool
    • Food Engineering Progress
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    • v.21 no.4
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    • pp.318-325
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    • 2017
  • Alcoholic steatosis is a fundamental metabolic disorder and may precede the onset of more severe forms of alcoholic liver disease. In this study, we isolated enzymatichydrolysate from Semisulcospira libertine by alcalase hydrolysis and investigated the protective effect of Semisulcospira libertine hydrolysate on liver injury induced by alcohol in the mouse model of chronic and binge ethanol feeding (NIAAA). In an in vitro study, the hydrolysate protects HepG2 cells from ethanol toxicity. Liver damage was assessed by histopathological examination, as well as by quantitating activities of serum aspartate aminotransferase (AST), alanine aminotransferase (ALT), and alkaline phosphatase (ALP). After the administration of S. libertina hydrolysate, fat accumulation and infiltration of inflammatory cells in liver tissues were significantly decreased in the NIAAA mouse model. The elevated levels of serum AST, ALT, and ALP activities, along with the lipid contents of a damaged liver, were recovered in experimental mice administrated with S. libertina hydrolysate, suggesting its role in blood enzyme activation and lipid content restoration within damaged liver tissues. Moreover, treatment with S. libertine hydrolysate reduced the expression rate of cyclooxygenase (COX-2), interleukin $(IL)-1{\beta}$, and IL-6, which accelerate inflammation and induces tissue damage. All data showed that S. libertine hydrolysate has a preventive role against alcohol-induced liver damages by improving the activities of blood enzymes and modulating the expression of inflammation factor, suggesting S. libertine hydrolysate could be a commercially potential material for the restoration of hepatotoxicity.

Effect of Fusion Procedure on the Development of Embryos Produced by Somatic Cell Nuclear Transfer in Hanwoo (Korean Cattle) (한우에서 융합방법이 체세포 핵이식 수정란의 발달에 미치는 영향)

  • Im, G.S.;Yang, B.S.;Park, S.J.;Chang, W.K.;Park, C.S.
    • Korean Journal of Animal Reproduction
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    • v.24 no.4
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    • pp.365-373
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    • 2000
  • The purpose of this study was to investigate the effects of the fusion pulses and fusion media on fusion rate and the development of embryos produced by somatic cell nuclear transfer in Hanwoo (Korean cattle). Nuclear donor cumulus and fetal fibroblast cells were cultured in Dulbecco's modified Eagle medium supplemented with 10% fetal bovine serum at 38.5$^{\circ}C$ in a humidified atmosphere of 5% $CO_2$in air. The in vitro matured oocytes were enucleated and then the isolated donor cells were introduced. The cumulus cell and cytoplast were fused using one pulse of 70 volts for 40$mutextrm{s}$, two pulses of 70 volts for 40$mutextrm{s}$ and one pulse of 180 volts for 15$mutextrm{s}$. The fetal fibroblast cell and cytoplast were fused using one pulse of 180 volts for 15$mutextrm{s}$ or 30$mutextrm{s}$. The cumulus cell and cytoplast were fused using mannitol and Zimmerman cell fusion medium (ZCFM) as a fusion medium. The fused embryos were activated after the fusion with 10 $\mu$M calcium ionophore for 5 min and 2 mM 6-dimethyl- aminopurine for 3 h. The nuclear transfer embryos were cultured in 500 ${mu}ell$ well of modified CR1aa supplemented with 3 mg/$m\ell$ BSA in th $\varepsilon$ four well dish cove red with mineral oil. After 3 days culture, culture medium was changed into modified CRlaa medium containing 1.5 mg/$m\ell$ BSA and 5% FBS for 4 days. The incubation environment was 5% $CO_2$, 5% $O_2$, 90% $N_2$ at 38.5$^{\circ}C$. When the cumulus cells were fused with enucleated oocytes by three different fusion pulses, one pulse of 180 volts for 15 $mutextrm{s}$ yielded the highest fusion rate and developmental rate to blastocyst among the pulses (P<0.05). When the fetal fibroblast cells were fused with enucleated oocytes, one pulse of 180 volts for 30$mutextrm{s}$ yielded significantly higher fusion rate compared with that for 15 $mutextrm{s}$(P<0.05). The present result indicates that the fusion rate between karyoplast and cytoplast was affected by the cell type and the optimal fusion condition was different according to cell type or size. When the fusion was conducted by the use of mannitol and ZCFM, the fusion rate was 71.2% and 65.8%, respectively. The developmental rates to blastocyst were 37.8% and 39.8%, respectively. There was no significant difference between two fusion media in the developmental rate of cumulus cell nuclear transfer embryos. These results indicate that optimal electric current should be selected according to cell type.

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Effect of Early Pregnant Heifer as Donor on the Ovum Pick-Up Derived Oocyte Aspiration and Embryo Production (초기 임신우의 공란우 활용이 초음파 유도 난자 채취 및 수정란 생산에 미치는 영향)

  • Jin, Jong-In;Kwon, Tae-Hyeon;Choi, Byeong-Hyun;Kim, Sung-Soo;Jo, Hyun-Tea;Bang, Jae-Il;Kim, Sam-Chul;Cho, Kyu-Woan;Lee, Jung-Gyu;Kong, Il-Keun
    • Journal of Embryo Transfer
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    • v.26 no.1
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    • pp.19-25
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    • 2011
  • This study was carried out to evaluate the effect of early pregnant cow as donor for Ovum Pick-Up (OPU) derived oocyte aspiration and embryo production in Holstein heifers. Four non-pregnant and 2 pregnant Holstein heifers were used as donor and then carried out total 17 OPU session for 10 weeks (2 times per week). Recovered cumulus-oocyte-complexes (COCs) were classified into 4 grade by oocyte cytoplasm and cumulus cells and matured in vitro in TCM-199 supplemented with 10% FBS, 10 mg/ml FSH and 1 mg/ml estradiol in 5% $CO_2$ and over 99% humidity for 24 h. After 24 h co-incubation with post-thaw sperm, the presumed zygotes were cultured in CR1aa medium with 4 mg/ml BSA for 3 days and then changed CR1aa medium with 10% of FBS for another 3~4 days. The Mean number of aspirated follicles and collected oocytes in the early stage pregnant and non-pregnant heifers were $13.0{\pm}4.3$ and $10.6{\pm}3.9$, $5.4{\pm}3.4$ and $7.7{\pm}3.6$ per session, respectively. Rate of collected oocyte from aspirated follicles were 59.2% and 50.5%, respectively. The average number of good quality oocytes (Grade I and II) in the early stage pregnant and non-pregnant heifers was $3.7{\pm}2.7$ and $4.9{\pm}2.6$ (Mean${\pm}$SD). Cleavage and blastocyst developmental rates in Grade I and II were 22.2% and 25.5%, and then $1.7{\pm}0.9$ and $1.4{\pm}1.1$ blastocyst per session, respectively. In conclusion, OPU technology can be used in early stage pregnant and non-pregnant heifers without any problem and so applied OPU derived embryo production to maximize the ability of genetically valuable females.