• Title/Summary/Keyword: Culture system

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Real-Time PCR for Quantitative Detection of Bovine Herpesvirus Type 1 (Bovine Herpesvirus Type 1 정량 검출을 위한 Real-Time PCR)

  • Lee, Dong-Hyuck;Jeong, Hyo-Sun;Lee, Jung-Hee;Kim, Tae-Eun;Lee, Jung-Suk;Kim, In-Seop
    • Korean Journal of Microbiology
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    • v.44 no.1
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    • pp.14-21
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    • 2008
  • Bovine blood, cell, tissue, and organ are used as raw materials for manufacturing biopharmaceuticals, tissue engineered products, and cell therapy. Manufacturing processes for the biologicals using bovine materials have the risk of viral contamination. Therefore viral validation is, essential in ensuring the safety of the products. Bovine herpesvirus type 1 (BHV-1) is the most common bovine pathogen found in bovine blood, cell, tissue, and organ. In order to establish the validation system for the BHV-1 safety of the products, a real-time PCR method was developed for quantitative detection of BHV-1 in raw materials, manufacturing processes, and final products as well as BHV-1 clearance validation. Specific primers for amplification of BHV-1 DNA was selected, and BHV-1 DNA was quantified by use of SYBR Green I. The sensitivity of the assay was calculated to be $2\;TCID_{50}/ml$. The real-time PCR method was validated to be reproducible and very specific to BHV-1. The established real-time PCR assay was successfully applied to the validation of Chinese hamster ovary (CHO) cell artificially infected with BHV-1. BHV-1 DNA could be quantified in CHO cell as well as culture supernatant. Also the real-time PCR assay could detect $10\;TCID_{50}/ml$ of BHV-1 artificially contaminated in bovine collagen. The overall results indicated that this rapid, specific, sensitive, and robust assay can be reliably used for quantitative detection of BHV-1 contamination during the manufacture of biologics.

In vitro Shoot Proliferation by Pulse Treatment from Shoot Cultures of Q. acutissima and Ex vitro Root Induction Using Peat Plug Systems in Quercus spp. (상수리나무 기내배양(器內培養)에서의 Pulse처리(處理)에 의한 줄기증식(增殖) 및 Peat Plug를 이용(利用)한 참나무류(類) 기내줄기의 기외삽목(器外揷木))

  • Moon, Heung Kyu;Youn, Yang;Son, Sung Ho;Lee, Suk Koo;Yi, Jae Seon
    • Journal of Korean Society of Forest Science
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    • v.82 no.3
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    • pp.221-226
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    • 1993
  • Methods for shoot proliferation via pulse treatment onto the microshoots of Quercus acutissima, and ex vitro root induction using peat plug systems of the microshoots of 4 oak trees were described. Pulsing solution was prepared by the addition of BA and/or BA plus zeatin onto the aqueous WPM and sterilized distilled water. Using the solution, pulsing time was adjusted at different levels(0. 1, 2, 5. 9, and 24 hours). Although the effect of pulsing solution prepared by the addition of cytokinins onto the sterilized distilled water was slightly lower in shoot proliferation rate, a little higher in shoot elongation was observed compared with that of aqueous WPM. One hour of pulse treatment revealed best in shoot proliferation and its elongation, whereas the increment of pulsing time slightly suppressed the response. In addition, prolonged pulse time resulted high frequency of hyperhydric shoot appearance. Single treatment of BA was better in shoot proliferation than that of BA combination with zeatin, whereas the latter treatment usually showed rapid and healthy shoot growth. For ex vitro root induction using peat plug systems, black oaks(Q. acutissima and Q. variabilis) revealed excellent rootability compared with white oaks(Q. serrata and Q. mongolica). Shoot-tip necrosis of white oaks eras one of the big problems for survival. In this study, we discribed the effect of pulse treatment, successful ex vitro rooting system by the incorporation of peat plug, and the possibilities for the overcoming the obstacles on micropropagation of oaks.

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A Study on Comics Outreach Programs for Contents marginalized Areas (콘텐츠 소외지역의 만화 아웃리치 프로그램 모델링 연구)

  • Lee, Seung-Jin
    • Cartoon and Animation Studies
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    • s.49
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    • pp.359-382
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    • 2017
  • Content is the complex of art and technology of trend, so it is important to experience different technologies for content education. Today, many non-profit organizations plan and operate numbers of programs for disabilities, low-income, and minority families to enhance the quality of life and the realization of social integration. These programs are limited to museums and galleries, not so pro-actively in progressing. Various contend education is necessary to the expansion of cultural exchange for the culturally alienated area. Naver is running an outreach program named . It is an experience-based outreach program where current cartoon / webtoon writers come directly to the school to inform students about the basic story of comics and comic techniques. However, the fact that the is not centered on the marginalized area but is centered on the Seoul Gyeonggi area, has the limitation that they can not benefit from a wide range of programs because they have a space limit of 'school', and, has a spatial limitation that the experience of the work is excluded. 'Outreach programs in marginalized areas' must be reorganized into a fluid dimension, not a fixed, single-system program. You should be able to experience and experience your work by directly using various professional equipment of comics based on your capacity and experience, local culture, religion, and society. These program participants will gain the effect of attractive and effective learning with empathy with their comic experience. Meanings of Comics content outreach program are following: First, the rich cultural archive can be used efficiently by providing various contents to existing outreach programs with the educational limitation of museums and galleries. Second, Comics contents can be enjoyed as a part of our life by understanding diversity and technology of contents. Third, because it is the program of expertise' participation, it can remodel, and restructure the severed experience in remote areas for the continuous growth and development, and furthermore, it can enhance the understanding of society.

$HgCl_2$ Dysregulates the Immune Response of Balb/c Mice (수은에 의한 마우스의 면역반응 조절장애)

  • Ki, No-Suk;Koh, Dai-Ha;Kim, Chong-Suh;Lee, Jung-Sang;Kim, Nam-Song;Lee, Hwang-Ho
    • Journal of Preventive Medicine and Public Health
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    • v.27 no.1 s.45
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    • pp.11-24
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    • 1994
  • The studies reported here were undertaken to investigate the effects of mercury chloride on immune system of Balb/c mouse employing a flexible tier of in vitro and in vivo assays. Mercury chloride inhibited the proliferative responses of spleen cells to lipopolysaccharide, pokeweed mitogen, and phytohemagglutinin as a dose-dependent manner. This inhibitory effect was observed not only when $HgCl_2$ was added 2nd or 3rd day of 3 days culture period but also when spleen cells was pretreated with $HgCl_2$ for 2 hours. Mercury chloride, however, potentiated the production of IgM and IgG from spleen cells. During the $HgCl_2$ administration by drinking for 3 weeks, the weight gain of mice was significantly blunted than that o control group mice, while no overt signs related to mercury toxicity were noted in any mice of experimental group. There was no change in thymus and spleen weights, and in histological findings of kidney, bone marrow of femur, thymus, spleen, and popliteal lymph node after 3 weeks of mercury exposure. However, $HgCl_2$ induced a significant increase of total serum IgM, IgG including $IgG_1,\;IgG_{2a}\;and\;IgG_{2b}$, and IgE in Balb/c mice. Treatment in vivo with anti-IL-4 monoclonal antibody significantly abrogated the $HgCl_2$-induced increase in total serum IgG1 and IgE. Whereas $HgCl_2$ potentiated total serum IgM and IgG, there was no difference in total serum hemagglutinin to SRBC (Sheep Red Blood Cell) between experimental and control group mice when these mice were immunized with SRBC. All these findings observed in Balb/c mice suggest that mercury perturbates well-orchestrated regulation of immune responses before developing histopathological changes in lymphoid tissues.

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Evaluation of Image Quality for Various Electronic Portal Imaging Devices in Radiation Therapy (방사선치료의 다양한 EPID 영상 질평가)

  • Son, Soon-Yong;Choi, Kwan-Woo;Kim, Jung-Min;Jeong, Hoi-Woun;Kwon, Kyung-Tae;Cho, Jeong-Hee;Lee, Jea-Hee;Jung, Jae-Yong;Kim, Ki-Won;Lee, Young-Ah;Son, Jin-Hyun;Min, Jung-Whan
    • Journal of radiological science and technology
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    • v.38 no.4
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    • pp.451-461
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    • 2015
  • In megavoltage (MV) radiotherapy, delivering the dose to the target volume is important while protecting the surrounding normal tissue. The purpose of this study was to evaluate the modulation transfer function (MTF), the noise power spectrum (NPS), and the detective quantum efficiency (DQE) using an edge block in megavoltage X-ray imaging (MVI). We used an edge block, which consists of tungsten with dimensions of 19 (thickness) ${\times}$ 10 (length) ${\times}$ 1 (width) $cm^3$ and measured the pre-sampling MTF at 6 MV energy. Various radiation therapy (RT) devices such as TrueBeam$^{TM}$ (Varian), BEAMVIEW$^{PLUS}$ (Siemens), iViewGT (Elekta) and Clinac$^{(R)}$iX (Varian) were used. As for MTF results, TrueBeam$^{TM}$(Varian) flattening filter free(FFF) showed the highest values of $0.46mm^{-1}$ and $1.40mm^{-1}$ for MTF 0.5 and 0.1. In NPS, iViewGT (Elekta) showed the lowest noise distribution. In DQE, iViewGT (Elekta) showed the best efficiency at a peak DQE and $1mm^{-1}DQE$ of 0.0026 and 0.00014, respectively. This study could be used not only for traditional QA imaging but also for quantitative MTF, NPS, and DQE measurement for development of an electronic portal imaging device (EPID).

Sensitivity of rice Plant to Potassium Stress of Various Growth Stages -II Effect of potassium depression on grain yield and its relation to nutrient content (생육시기별수도(生育時期別水稻)의 가리부족(加里不足)에 대(對)한 감수성(感受性) -II 수량(收量)에 대(對)한 가리결제(加里缺除)의 영향(影響) 및 수량(收量)과 양분함량(養分含量)과의 관계(關係))

  • Park, Hoon;Mok, Sung Kyun;Kim, Sung Bae
    • Korean Journal of Soil Science and Fertilizer
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    • v.7 no.3
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    • pp.163-175
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    • 1974
  • Effects on yields, yield components and nutrient content of potassium depression for two or three weeks at various growth stages were investigated in rice (var. Jinheung) under sand culture system.(K 40 ppm 1973) 1. Analysis of variance showed significant difference among treatments of both two-week (at p=0.01) and three-week depression (at p=0.05) in yield. 2. Most sensitive stage to potassium depression on yield appeared two weeks until heading (42% yield decrease) and sensitivity decreased the growth stage is apart from heading either before or after. During 30 days after transplanting two-week potassium depression increased yield, but three-week depression decreased yield. Until about 30 days after heading depression caused poor yield. 3. Root potassium involves in harvest index, filled grain ratio and grain weight with significant correlation and considerably in spikelet per panicle while potassium in leaf sheath+culm involves considerably in spikelet per panicle and panicle per hill. Relative total dry matter weight was significantly correlated with panicle per hill, spikelet per panicle and K or K/Ca+Mg only in leaf sheath+culm. The indications are that root potassium contributes for building sink and efficiency of structure while potassium in leaf sheat+culm primarily for building source, productive structure. 4. Relative yield was significantly correlated with potassium content in root and leaf sheath+culm and with K/Ca+Mg and its ratio before and after depression in root indicating that potassium depression occurs greatly in root and that K/Ca+Mg might have more important role than K content alone under depression. 5. Optimum level of $K_2O$ appears around 3% in leaf blade. 4% in leaf sheath+culm and 1% in root under the assumption that below these level the same content has the same role in relation to yield during growth. The K/Ca+Mg appeares to be 2.5 in root and should not decrease throughout the growth stages. 6. The increase of sodium content in plant by K depression was highest, especially in leaf sheath during the most insensitive period to K depression suggesting that insensitivity may be attributed to partial replacement of Na for K. Partial replacement seems very little in sensitive stage (later stage) and sensitive organ (root).

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Identification of Antagonistic Bacteria, Pseudomonas aurantiaca YC4963 to Colletotri­chum orbiculare Causing Anthracnose of Cucumber and Production of the Antibiotic Phenazine-l-carboxylic acid (Colletotrichum orbiculare에 대한 길항세균 Pseudomonas aurantiaca YC4963의 분리 동정 및 항균물질 Phenazine-1-carboxylic acid의 생산)

  • Chae Hee-Jung;Kim Rumi;Moon Surk-Sik;Ahn Jong-Woong;Chung Young-Ryun
    • Korean Journal of Microbiology
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    • v.40 no.4
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    • pp.342-347
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    • 2004
  • A bacterial strain YC4963 with antifungal activity against Colletotrichum orbiculare, a causal organism of cucumber anthracnose was isolated from the rhizosphere soil of Siegesbeckia pubescens Makino in Korea. Based on physiological and biochemical characteristics and 16S ribosomal DNA sequence analysis, the bac­terial strain was identified as Pseudomonas aurantiaca. The bacteria also inhibited mycelial growth of several plant fungal pathogens such as Botrytis cinerea, Fusarium oxysporum and Rhizoctonia solani on PDA and 0.1 TSA media. The antifungal activity was found from the culture filtrate of this isolate and the active compound was quantitatively bound to XAD adsorption resin. The antibiotic compound was purified and identified as phenazine-l-carboxylic acid on the basis of combined spectral and chemical analyses data. This is the first report on the production of phenazine-l-carboxylic acid by Pseudomonas aurantiaca.

Effects of Manipulation Conditions on Development of Nuclear Transplant Bovine Embryos Derived from In Vitro Matured Oocytes (미세조작조건이 소 핵이식배의 발달에 미치는 영향)

  • 최상용;노규진;공일근;송상현;조성근;박준규;이효종;박충생
    • Korean Journal of Animal Reproduction
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    • v.21 no.3
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    • pp.293-302
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    • 1997
  • Follicular oocytes of Grade I and II were collected from 2~6 mm ovarian follicles and matured in vitro (IVM) for 24 hrs in TCM-199 su, pp.emented with 35$\mu\textrm{g}$/ml FSH, 10$\mu\textrm{g}$/ml LH, and 1$\mu\textrm{g}$/ml estradiol-17$\beta$ at 39$^{\circ}C$ under 5% CO2 in air. They were fretilized in vitro (IVF) by epididymal spermatozoa capacitated with heparin for 12 hrs. The zygotes were then co-cultured in vitro with bovine oviducted epithelial cells (BOEC) for 7 to 9 days. The optimal time for IVM, the successful enucleation of IVM oocytes by micromanipulation at different oocyte ages after IVM, and the ideal culture system for IVM for effective IVF and in vitro development of IVM-IVF embryos was examined for in vitro production of nuclear recipient oocytes and nuclear donor embryos. To improve the efficiency of nuclear transplantation (NT) of IVF embryo into IVM follicular oocytes, this study evaluated the optimal electric condition and oocytes age for activation of IVM oocytes and in vitro development of NT embryos. In vitro development of NT embryos with preactivation or non-preactivation in enucleation oocytes, cell number of IVN-IVF embryos, and NT embryos wre also examined. The results obtained were as follows; 1. The most suitable enucleation time was at 24 hpm (83.3%) rather than that of 28 hpm(69.6%) and 32 hpm(50.0%). 2. There was no difference among the fusion rates of NT embryos at the voltages of 0.75, 1.0 and 1.5 kV/cm, but the in vitro development rates to morule and blastocyst were significantly (P<0.05) higher at the voltage of 0.75(12.5%) and 1.0kV/cm (12.6%) compared to 1.5kV/cm(0%). 3. No significant difference in activation rates were seen in NT embryos stimulated for 30, 60 and 120 $\mu$sec (71.7, 85.2 and 71.9%, respectively), but the in vitro development rates to morulae and blastocyst were significantly (P<0.05) higher in the oocytes stimulated for 30 $\mu$sec (11.6%) and 60 $\mu$sec(10.7%) than 120 $\mu$sec(0.0%). 4. The fusion rates (71.0 and 87.3%) and the in vitro development rates (9.1 and 12.7%) to morula and blastocyst were seen in the NT embryos stimulated at 28 and 32 hpm under the condition of 1.0 kV/ml, 60 $\mu$sec. However, at 24 hpm the fusion rates were 64.8% and the in vitro development to morula and blastocyst were not seen. 5. The fusion rates between the 8~12, 13~17 and 18~22-cell stage of IVM-IVF embryos were not significantly different. The in vitro development rates of the fused embryos to morula and blastocyst which were received from a blastomere of 8~12, 13~17 and 18~22-cell stages of IVM-IVF embryos were 14.9, 8.3 and 6.5%, respectively. 6. The in vitro development rate of the enucleated recipient oocytes with preactivation (24.2%) to morula and blastocyst was significantly (P<0.05) higher than that of non-preactivation (12.8%). 7. The cell numbers of NT blastocyst and IVM-IVF blastocyst cultured during 7~9 days were 63$\pm$11 and 119$\pm$23, and then their the mean cell cycle number were 5.98 and 6.89, respectively.

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Trend and Further Research of Rice Quality Evaluation (쌀의 품질평가 현황과 금후 연구방향)

  • Son, Jong-Rok;Kim, Jae-Hyun;Lee, Jung-Il;Youn, Young-Hwan;Kim, Jae-Kyu;Hwang, Hung-Goo;Moon, Hun-Pal
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.47
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    • pp.33-54
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    • 2002
  • Rice quality is much dependent on the pre-and post harvest management. There are many parameters which influence rice or cooked rice qualitys such as cultivars, climate, soil, harvest time, drying, milling, storage, safety, nutritive value, taste, marketing, eating, cooking conditions, and each nations' food culture. Thus, vice evaluation might not be carried out by only some parameters. Physicochemical evaluation of rice deals with amy-lose content, gelatinizing property, and its relation with taste. The amylose content of good vice in Korea is defined at 17 to 20%. Other parameters considered are as follows; ratio of protein body-1 per total protein amount in relation to taste, and oleic/linoleic acid ratio in relation to storage safety. The rice higher Mg/K ratio is considered as high quality. The optimum value is over 1.5 to 1.6. It was reported that the contents of oligosaccharide, glutamic acid or its derivatives and its proportionalities have high corelation with the taste of rice. Major aromatic compounds in rice have been known as hexanal, acetone, pentanal, butanal, octanal, and heptanal. Recently, it was found that muco-polysaccharides are solubilized during cooking. Cooked rice surface is coated by the muco-polysaccharide. The muco-polysaccharide aye contributing to the consistency and collecting free amino acids and vitamins. Thus, these parameters might be regarded as important items for quality and taste evaluation of rice. Ingredients of rice related with the taste are not confined to the total rice grain. In the internal kernel, starch is main component but nitrogen and mineral compounds are localized at the external kernel. The ingredients related with taste are contained in 91 to 86% part of the outside kernel. For safety that is considered an important evaluation item of rice quality, each residual tolerance limit for agricultural chemicals must be adopted in our country. During drying, rice quality can decline by the reasons of high drying temperature, overdrying, and rapid drying. These result in cracked grain or decolored kernel. Intrinsic enzymes react partially during the rice storage. Because of these enzymes, starch, lipid, or protein can be slowly degraded, resulting in the decline of appearance quality, occurrence of aging aroma, and increased hardness of cooked rice. Milling conditions concerned with quality are paddy quality, milling method, and milling machines. To produce high quality rice, head rice must contain over three fourths of the normal rice kernels, and broken, damaged, colored, and immature kernels must be eliminated. In addition to milling equipment, color sorter and length grader must be installed for the production of such rice. Head rice was examined using the 45 brand rices circulating in Korea, Japan, America, Australia, and China. It was found that the head rice rate of brand rice in our country was approximately 57.4% and 80-86% in foreign countries. In order to develop a rice quality evaluation system, evaluation of technics must be further developed : more detailed measure of qualities, search for taste-related components, creation and grade classification of quality evaluation factors at each management stage of treatment after harvest, evaluation of rice as food material as well as for rice cooking, and method development for simple evaluation and establishment of equation for palatability. On policy concerns, the following must be conducted : development of price discrimination in conformity to rice cultivar and grade under the basis of quality evaluation method, fixation of head rice branding, and introduction of low temperature circulation.

Differentiation and Apoptosis of the Mammalian Embryo and Embryonic Stem Cells(ESC): I. Establishment of Mouse ESC and Induction of Differentiation by Reproductive Hormones (포유동물의 배아 및 기간세포의 분화와 세포사멸 기작: I. 생쥐 배아줄기세포의 확립과 분화유도에 미치는 생식호르몬의 영향)

  • 성지혜;윤현수;이종수;김철근;김문규;윤용달
    • Development and Reproduction
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    • v.6 no.1
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    • pp.55-66
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    • 2002
  • Embryonic stem cells(ES cells) are derived from the inner cell mass(ICM) of blastocysts, which have the potentials to remain undifferentiated, to proliferate indefinitely in vitro, to differentiate into the derivates of three embryonic germ layers. ES cells are an attractive model system for studying the initial developmental decisions and their molecular mechanisms during embryogenesis. Additionally, ES cells of significant interest to those characterizing the various gene functions utilizing transgenic and gene targeting techniques. We investigated the effects of reproductive hormones, gonadotropins(GTH) and steroids on the induction of differentiation and expressions of their receptor genes using the newly established mouse ES cells. We collected the matured blastocysts of inbred mice C57BL/6J after superovulation and co-cultured with mitotically inactivated STO feeder cells. After 5 passages, we confirmed the expression alkaline phosphatase(Alk P) activity and SSEA-1, 3, 4 expressions. The protocol devised for inducing ES differentiation consisted of an aggregation steps, after 5 days as EBs in hormone treatments(FSH, LH, E$_2$, P$_4$, T) that allows complex signaling to occur between the cells and a dissociation step, induced differentiation through attachment culture during 7 days in hormone treatments. Hormone receptors were not increased in dose-dependent manner. All hormone receptors in ES cells treated reproductive hormones were expressed lower than those of undifferentiated ES cell except for LHR expression in E$_2$-treated ES cells group. After hormone induced differentiation, at least some of the cells are not terminally differentiated, as is evident from the expression of Oct-4, a marker of undifferentiated. To assess their differentiation by gene expression, we analyzed the expression of 7 tissue-specific markers from all three germ layers. Most of hormone-treated group increased in the expression of gata-4 and $\alpha$ -fetoprotein, suggesting reproductive hormone allowed or induced differentiation of endoderm.

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