• 제목/요약/키워드: Co-culture system

검색결과 472건 처리시간 0.03초

우 난포란의 체외수정과 발육 (In Vitro Fertilization and Development of Bovine Oocytes)

  • 김정익
    • 한국가축번식학회지
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    • 제13권2호
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    • pp.98-104
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    • 1989
  • Successful techniques of in vitro fertilization(IVF) are valuable for studying the process of fertilization and for developing economical procedures for gene and nuclear transfer in farm animals. To date, bovine IVF system has been developed with oocytes in vitro or vitro, but the resulting zygotes exhibit limited embryonic development after in vitro culture. Even though in vitro matured oocytes achieved high fertilization and cleavage rates, these embryos appear extremly low rate of pregnancies when transferred to synchronized recipients. Development of early bovine embryos in vitro is generally arrested at the 8-to 16-cell stage. However, recent use of somatic cells such as trophoblastic vesicle, granulosa and oviduct epithelial cell for co-culture with early bovine embryos has proven effective for development of embryos, matured and fertilized in vitro, past the in vitro cell blocks. These factors clearly indicate the value of the co-culture system in promoting development of bovine oocytes matured and fertilized in vitro to morula or blastocyst stage in vitro. In addition, co-culture system may beome a tool for evaluation of viability of ova that have been manipulated by procedures such as splitting, microinjection and nuclear transfer.

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돼지 자궁내막 상피세포와 공동배양된 Collagen Matrix Gel을 이용한 체외수정란 배양체계 확립 (Establishment of Embryo Culture System using Co-incubated Collagen Matrix Gel with Porcine Endometrial Cells)

  • 이상희;한혜인;황보용;이승형;정희태;양부근;박춘근
    • Reproductive and Developmental Biology
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    • 제39권3호
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    • pp.49-57
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    • 2015
  • In order to achieve successful in vitro production of embryo, it is necessary to establish intrauterine environment during in vitro culture. Thus, this study was investigated to establish embryo culture system using co-incubated collagen matrix gel (CM) with endometrial epithelial cells (EC). Endometrial epithelial cells were isolated from porcine endometrium at follicular phase, the cells seeded in insert dish for co-incubation with CM-coated culture dish. Then, culture media treated with/without 2.0 IU/ml hCG or 10 ng/ml $IL-1{\beta}$. After incubation for 24 h, the co-incubated insert dishes were removed from CM-coated culture dish before embryo culture. Embryos at 48 h after in vitro fertilization (IVF) were cultured on the dish for 120 h with porcine zygote medium. We determined PTGS-2 expression in the ECs, VEGF protein in co-incubated CM with EC and observed cleavage rate and blastocyst development of embryos at 168 h after IVF. In result, expression of PTGS-2 was higher at co-incubated EC with hCG and $IL-1{\beta}$ groups than EC without hCG and $IL-1{\beta}$. The VEGF protein was detected at co-incubated CM with EC, EC treated with hCG and $IL-1{\beta}$ groups higher than CM group. Also, cleavage rate was no significantly difference among all group, however, blastocyst development was significantly higher in co-incubated CM with EC treated with hCG group than un-treated groups (p<0.05). Therefore, we suggest that novel embryo culture system using co-incubated collagen matrix gel with endometrial epithelial cells treated with $IL-1{\beta}$ is beneficial and useful for enhancing the production of porcine blastocysts in vitro.

공배양 및 산소농도가 한우 난포란의 체외발생에 미치는 영향 (The Effect of Co-culture and Oxygen Concentration on In Virto Fertilization of Follicular Oocytes in Korean Native Cattle)

  • 이재관;윤준진;황성수;윤종택;김창근;정영채
    • 한국가축번식학회지
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    • 제22권1호
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    • pp.43-50
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    • 1998
  • The effect of oxygen tension on embryonic development in co-culture was evaluated from the standpoint of the reduction of dissolved oxygen concentration by the oxygen consumption of feeder cells. Three co-culture systems using bovine oviductal epitherial cells (BOEC), African green monkey kidney cells (Vero cells) or buffalo rat liver cells (BRLC) have been compared in terms of development of bovine embryos derived from oocytes matured and fertilized in vitro. Among the co-cultured embryo, Vero cells su, pp.rted the highest developmental rate (29%) and the other two showed the similar rates. When the co-cultures were incubated in three different oxygen tension such as 5, 10, 20% oxygen atmosphere, embryos co-cultured with Vero cells at 10%-O2 resulted in the highest percentage of development. From the measurement of oxygen consumption of feeder cells, BRLC consumed 1.38 10-10 mg-O2/min/cell which was higher than 0.94 10-10 and 0.26 10-10mg-O2/min/cell for Vero cells and BOEC, respectively. Based on the oxygen consumption data, the phenomena of optimum oxygen tension required in embryo development in vitro has been analyzed, and we suggested that gas phase oxygen concentration, oxygen consumption rate of feeder cells and the number of feeder cells should be considered for the design of optimal co-culture system for effective fertilization of embryos in vitro.

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Protective Effect of BOEC Co-Culture System against Nitric Oxide on Development of Bovine IVM/IVF Embryos

  • Jang, Hyun-Yong;Jung, Yu-Sung;Li, Zheng-Yi;Yoon, Hyoung-Jong;Cheong, Hee-Tae;Kim, Jong-Taek;Park, Choon-Keun;Yang, Boo-Keun
    • Reproductive and Developmental Biology
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    • 제32권3호
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    • pp.167-173
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    • 2008
  • Somatic cells such as oviduct epithelial cell, uterine epithelial cell, cumulus-granulosa cell and buffalo rat river cell has been used to establish an effective culture system for bovine embryos produced in in vitro. But nitric oxide (NO) metabolites secreted from somatic cells were largely arrested the development of bovine in vitro matured/ in vitro fertilized (IVM/IVF) embryos, suggesting that NO was induced the embryonic toxic substance into culture medium. The objective of this study was to investigate whether BOEC co-culture system can ameliorate the NO-mediated oxidative stress in the culture of bovine IVM/IVF embryos. Therefore, we evaluated the developmental rate of bovine IVM/IVF embryos under BOEC co-culture system in the presence or absence of sodium nitroprusside (SNP), as a NO donor, and also detected the expression of growth factor (TGF-$\beta$, EGF and IGFBP) and apoptosis (Caspase-3, Bax and Bcl-2) genes. The supplement of SNP over 5 uM was strongly inhibited blastocyst development of bovine IVM/IVF embryos than in control and 1 uM SNP group (Table 2). The developmental rates beyond morulae stages of bovine IVM/IVF embryos co-cultured with BOEC regardless of SNP supplement (40.4% in 5 uM SNP+ BOEC group and 65.1% in BOEC group) were significantly increased than those of control (35.0%) and SNP single treatment group (23.3%, p<0.05: Table 3). The transcripts of Bax and Caspase-3 genes were detected in all experiment groups (1:Isolated fresh cell (IFC), 2:Primary culture cell (PCC), 3:PCC after using the embryo culture, 4: PCC containing 5 uM SNP and 5: PCC containing 5 uM SNP after using the embryo culture), but Bcl-2 gene was not detected in IFC and PCC (Fig. 1). In the expression of growth factor genes, TGF-$\beta$ gene was found in all experimental groups, and EGF and IGFBP genes were not found in IFC and PCC (Fig. 2). These results indicate that BOEC co-culture system can increase the development beyond morula stages of bovine IVM/IVF embryos, possibly suggesting the alleviation of embryonic toxic substance like nitric oxide.

한우 체외수정란의 체외배양, 동결보존 및 이식에 관한 연구 I. 한우 체외수정란의 체외배양에 대한 공배양세포와 성장인자의 효과 (Studies on In Vitro Culture, Freezing and Transfer of Korean Native Cattle Embryos Fertilized In Vitro I. Effect of Co-culture Cells and Growth Factors on In Vitro Development of Korean Native Cattle Embryos Fertilized In Vitro)

  • 김일화;손동수;이호준;최선호;양병철;이광원;김경남;장인호
    • 한국수정란이식학회지
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    • 제11권2호
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    • pp.111-124
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    • 1996
  • The present study was carried out to investigate the effects of co-culture cells and growth factors on in vitro culture of Korean native cattle(KNC) embryos fertilized in vitro. Two-eight cell embryos were cultured in vitro using 4 types of co-culture cells and 3 growth factors singly or in combination. The results were as follows, In the co-culture of 2~8 cell embryos with bovine oviductal epithelial cell(BOEC), granulosa cell(BGC), uterine epithelial cell(BUEC) and mouse embryonic fibroblast (MEF) monolayers, the developing rate to blastocysts was significantly(P<0.05) higher with BUEC(32.1%) than with MEF(15.3%), BGC(13.2%) and non co-culture control(11.6%). When the morula co-cultured with BOEC for 5 days following in vitro fertilization were co-cultured with BOEC continuously or with BUEC, respectively, the developing rate to blastocysts was higher with BUEC(73.9%) than with BOEC(56.0%). To examine the effects of growth factors on in vitro development of 2~8 cell embryos, epidermal growth factor(EGF), transforming growth factor-$\beta$l(TGF-$\beta$l) and insulin-like growth factor-1(IGF-1) were added singly or in combination to TCM 199 maturation medium with respective concentration. In a addition of each 10, 30 and SOng /rnl EGF, the developing rate to blastocysts was the highest in lOng /ml EGF(25.3%). In addition of each 1, 2 and Sng /mi TGF-$\beta$1, the developing rate to blastocysts was the highest in lng /ml TGF-$\beta$1(28.8%). In addition of each 50, 100ng/ml JGF-l, the developing rate to blastocysts was higher in 100ng/ml IGF-l(16.5%) than in SOng/mi IGF-1(12.9%). When lOng /ml EGF and lng /ml TGF-$\beta$l was added singly or in combination, the developing rate to blastocysts was similar in groups added singly or in combination with EGF and TGF-$\beta$l (23.l~24.6%), although higher than in control(16.7%). In the co-culture of 2~8 cell embryos Wth BOEC + each 10, 30 and 5Ong /rnl EGF, the developing rate to blastocysts was significantly(p<0.05) higher in BOEC + long /ml EGF(32.3%) than in BOEC + 3Ong /ml EGF(18.9%) and BOEC + song /ml EGF(9.7%). In the co-culture of 2~8 cell embryos with BOEC + each 1, 2, Sng /ml TGF-$\beta$l the developing rate to blastocysts was higher in BOEC + Sng/rnl TGF-$\beta$l(28.2%) than in BOEC + lng /ml TGF-$\beta$l(21.7%) and BOEC + 2ng/ml TGF-$\beta$l(21.4%). In summary, higher developing rate to blastocysts were obtained with co-culture of BUEC for co-culture system, with addition of lOng /ml EGF or lng /ml TGF-$\beta$l for growth factor culture system, and with co-culture of BOEC + lOng /ml EGF or BOEC + Sng /ml TGF-$\beta$l for co-culture + growth factor culture system.

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Aspergillus awamori와 Zymomonas mobilis로 구성된 혼합고정화 배양계의 최적 조건 (Optimal Conditions of Co-Immobilized Mixed Culture System with Aspergillus awamori and Zymomonas mobilis)

  • 박석규;이상원;손봉수;최수철;서권일;성낙계;김홍출
    • 한국식품영양과학회지
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    • 제24권5호
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    • pp.803-810
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    • 1995
  • Gel bead의 효율적인 이용을 목적으로 산소요구성이 전혀 다른 두 균주로 구성된 혼합고정화 배양계의 개발을 시도하였다. 호기성 균으로서 A. awamori, 혐기성균으로서 Z. mobilis를 사용하여 생전분으로 부터 에탄올 생산을 행한 결과는 다음과 같다. 두 균주의 최적 혼합비율은 A. awamori $1.25{\times}10^{9}\;spore/L-gel$, Z. mobilis 0.5g cell/L-gel이었다. 배양이 완료된 후의 gel bead 내의 균주 분포는 각각 혐기부와 호기부, 즉 gel bead의 표면부와 중심부로 생육장소가 구분되어 있었다. A-Z계의 배양에서는 에탄올의 수율이 낮았고, 균사가 gel bead로 부터 누출되었다. 배양 개시 후 36시간째에 배양기의 면전을 특수 제조한 check valve가 부착된 실리콘 plug로 교환하여 혐기적 배양을 행한 A-Z 36계에서는 gel bead로 부터 균사의 누출이 상당히 억제되었고, pH가 4.3을 유지하면서, 에탄올의 수율이 대조구 보다 약2배 높게 나타났다.

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Rhizopus japonicus와 Zymomonas mobilis의 혼합고정화 배양계에 의한 생전분으로부터 에탄올 생산 (Ethanol Production from Raw Starch by Co-Immobilized Mixed Rhizopus japonicus and zymomonas mobilis)

  • 최수철;이상원;박석규;성찬기;손봉수;성낙계
    • 한국식품영양과학회지
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    • 제25권4호
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    • pp.708-714
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    • 1996
  • 호기성의 Rhizopus japonicus와 혐기성 Zymomonas mobilis로 구성된 혼합고정화 배양계(R-Z계)를 제조하고, 생전분으로부터 에탄올 생산에 응용하였다. R. japonicus를 고정화배양하므로서 액체배양에서 보다 2배 높은 glucose량을 얻었다. R-Z계의 에탄올 생산량은 1.67g/L(Yp/s, 0.094)이 었지만, 배양 24시간째부터 산소공급을 억제한 R-Z 24계에서는 6.54g/L(Yp/s, 0.38)의 에탄올을 얻어 대조구의 약 4배를 향상시켰다. 회분배양에서는 5%의 기질 농도가 적당하였으며, 생산된 에탄올은 15.02g/L(Yp/s, 0.36)이었다. 2% 기질을 5회 첨가한 유가배양에서는 2%기질의 회분배양과 동등한 수율인 0.38을 얻었다.

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최적 배양 조건을 이용한 CO2 제거 목적의 담수 미세조류 Parachlorella kessleri의 바이오매스 생산성 향상 (Enhanced Biomass Productivity of Freshwater microalga, Parachlorella kessleri for Fixation of Atmospheric CO2 Using Optimal Culture Conditions)

  • 김지훈;홍선우;김진우;손병락;김미경;김용환;설진현;전수환
    • 한국해양바이오학회지
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    • 제16권1호
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    • pp.36-44
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    • 2024
  • This study attempted to improve the growth of the freshwater microalgae, Parachlorella kessleri, through the sequential optimization of culture conditions. This attempt aimed to enhance the microalgae's ability to fixate atmospheric CO2. Culture temperature and light intensity appropriate for microalgal growth were scanned using a high-throughput photobioreactor system. The supplied air flow rate varied from 0.05 to 0.3 vvm, and its effect on the growth rate of P. kessleri was determined. Next, sodium phosphate buffer was added to the culture medium (BG11) to enhance CO2 fixation by increasing the availability of CO2(HCO3-) in the culture medium. The results indicated that optimal culture temperature and light intensity were 20℃-25℃ and 300 μE/m2/s, respectively. Growth rates of P. kessleri under various air flow rates highly depended on the increase of the culture's flow rate and pH which determines CO2 availability. Adding sodium phosphate buffer to BG11 to maintain a constant neutral pH (7.0) improved microalgal growth compared to control conditions (BG11 without sodium phosphate). These results indicate that the CO2 fixation rate in the air could be enhanced via the sequential optimization of microalgal culture conditions.

한우 난포란 유래 배반포의 체외생산을 위한 생물학적 요인들의 영향 (Effects of biological Factors on In Vitro Production of Hanwoo Embryos)

  • 박흠대;김재영;주재홍;공건오;윤산현;공일근;이상민;이상진;송해범
    • 한국수정란이식학회지
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    • 제15권2호
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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.

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Cody Recommendation System Using Deep Learning and User Preferences

  • Kwak, Naejoung;Kim, Doyun;kim, Minho;kim, Jongseo;Myung, Sangha;Yoon, Youngbin;Choi, Jihye
    • International Journal of Advanced Culture Technology
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    • 제7권4호
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    • pp.321-326
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    • 2019
  • As AI technology is recently introduced into various fields, it is being applied to the fashion field. This paper proposes a system for recommending cody clothes suitable for a user's selected clothes. The proposed system consists of user app, cody recommendation module, and server interworking of each module and managing database data. Cody recommendation system classifies clothing images into 80 categories composed of feature combinations, selects multiple representative reference images for each category, and selects 3 full body cordy images for each representative reference image. Cody images of the representative reference image were determined by analyzing the user's preference using Google survey app. The proposed algorithm classifies categories the clothing image selected by the user into a category, recognizes the most similar image among the classification category reference images, and transmits the linked cody images to the user's app. The proposed system uses the ResNet-50 model to categorize the input image and measures similarity using ORB and HOG features to select a reference image in the category. We test the proposed algorithm in the Android app, and the result shows that the recommended system runs well.