• Title, Summary, Keyword: Medium development

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Effect of Conditioned Medium of Human Endothelial Cell Line(tHUE-2 cell) on In Vitro Development of Mouse 1-cell Embryos In Vitro Fertilized (체내 수정된 Mouse 1-세포배의 체외발생에 미치는 혈관내피세포주(tHUE-2세포) 배양액의 영향)

  • ;;;Y. Mitsui
    • Korean Journal of Animal Reproduction
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    • v.19 no.2
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    • pp.81-88
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    • 1995
  • Culture medium (ASF-301) of tHUE-2 cell, human endothelial cell line, and culture medium of these cells (conditioned medium : CM) which affect embryonic development of in vivo fertilized 1-cell embryos of mouse were examined. Two-cell stage block of mouse embryos was overicomed in ASF-301 and CM without EDTA, which usually added in basic medium (modified Whitten Medium: MWM, control) to overcome the 2-cell stage block. The developmental rates of embryos to the blastocyst stage were significantly increased in MWM containing 12.5% of growth factors added to ASF-301 (10mg/ $\ell$ transferrin, 1mg/$\ell$ insulin, 0.01mg/$\ell$ EGF) than those of 100% addition and control, 78.0% vs 20.8 and 52.3%, respectively (P<0.05), but the growth factors was not affected the hatching rate of blastocyst. Using ASF-301 or CM which was not treated, embryonic development into the blastocyst and hatched blastocyst stages were not affected. However, proportions of embryonic development into the blastocyst and hatched blastocyst stages were significantly higher in dilution (ASF-301 1:10; CM 1:3~1:6) than those in control (P,0.05). In ASF-301 dialyzed M.W.<10000 dialysis membrane, the developmental rate upto the hatched blastocyst stage was significantly increased, compared to ASF-301 which was not dialyzed (P<0.05), and hatching rate of blastocyst of these group was singnificantly increased than those in MWM (P<0.05). Compared to CM which was not dialyzed, however, in dialyzed CM was significantly decreased, compared to untreated CM (P<0.05), especially any hatched blastocyst was not appeared. As a result of these experiments indicated that a kind or porper treatment such as a dilution of complex synthetic cell culture medium and conditioned medium, and that a optimal concentration of growth factors are usuful for embryo cultrue in vitro.

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Effects of established cell lines on bovine embryo development during in vitro culture (계대세포를 이용한 소 수정란의 체외배양 체계에 관한 연구)

  • Joo, Suk-chun;Lee, Byeong-chun;Lee, Won-yu;Choi, Yun-seok;Hwang, Woo-suk
    • Korean Journal of Veterinary Research
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    • v.37 no.3
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    • pp.647-659
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    • 1997
  • To overcome the difficulties of collecting and culture of primary cell from genital tract on embryonic development, the present study was carried out to investigate the critical effect of cell lines, such as BRL and Vero cell and its conditioned medium on the development of early Korean native cattle embryos in vitro. Oocytes collected from slaughterhouse ovaries were matured in TCM199 containing FSH, estradiol-$17{\beta}$ and FBS with granulosa cell monolayer for 24 hours and then fertilized in vitro using frozen-thawed, heparin-treated spermatozoa in TALP for 30 hours. And then early embryos (1-2cell) were cultured in TCM199 containing 10% FBS with BOEC, Granulosa, BRL, Vera cell monolayers and conditioned medium for 2~3 days. Development to morulae and blastocysts were recorded, also examined the number of blastomeres presented a valuable parameter for the evaluation of embryonic development. The early cleavage rates of in vitro fertilized embryos co-cultured, there was no differences between primary cell and cell lines(p<0.05). The rate of development to the later stage, coculture of BRL cell was significantly higher than that of the primary cell(p<0.05). The rates of development to morula and blastocyst were significantly higher in vero cell than BRL, Granulosa, Oviduct epithelial cell conditioned medium. In the result of effect of serum on development of early bovine embryos, the use of media containing serum were significantly higher than the use of not containing one on development of early and later stage of embryos. The result of number of blastomeres in blastocysts, there is no differences between primary cell and cell lines. The blastocysts from coculture were higher than from conditioned medium in blastomere cells. In summary, these experments have proved that the culture system in TCM199 with BRL, Vero cell monolayers is effective on in vitro development of early bovine embryos, In addition, it is effective to development of bovine embryos that containing serum in conditioned medium, or in co-culture rather than in conditioned medium alone. The use of cell lines opponent to primary cells is effective in bovine embryo culture.

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Optimization of Embryo Density and the Volume of Culture Medium for an Improvement of Mouse Parthenogenetic Embryo Development

  • Roh Sangho;Choi Young-Joo;Min Byung-Moo
    • 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.

Effect of Essential and Non-essential Amino Acids, Taurine or Glycine Supplemented to Fertilization Medium on In Vitro Cleavage and Development of Bovine Oocytes Matured and Fertilized In Vitro (체외수정배양액내에 첨가된 필수 .비필수아미노산, Taurine 또는 Glycine이 소 난자의 분할 및 체외발육에 미치는 영향)

  • 이은송;이병천;황우석
    • Journal of Embryo Transfer
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    • v.10 no.2
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    • pp.131-138
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    • 1995
  • Essential and non-essential amino acids supplemented to culture medium stimulate mammalian embryo development in vitro. Amino acids such as glycine, taurine and alanine are concentrated in the lumen of oviduct and uterus and it can he thought that these amino acids may have physiological role on fertilization and embryo development. Our aim of this experiment was to investigate the effects of essential and non-essential amino acids, taurine or glycine supplemented to fertilization medium on the cleavage and subsequent in vitro development of bovine oocytes matured and fertilized in vitro. Immature oocytes were obtained from slaughtered Holstein cows and heifers and matured in TCM199 containing 10% fetal calf serum, 2.5 $\mu$g /mL of FSH and LH and 1 $\mu$g / mL of estradiol with granulosa cells in vitro. After maturation, oocytes were coincubated with sperm in fertilization medium supplemented with Minimum Essential Medium (MEM) essential and non-essential amino acids, taurine (3.75 mM) or glycine (10 mM) for 30 hours in vitro. Inseminated oocytes were cultured in synthetic oviduct fluid medium (SOEM) containing MEM essential, non-essential amino acids and 1 mM glutarnine up to 8 days after fertilization.Supplementation of fertilization medium with MEM essential and non-essential amino acids lowered significantly (p<0.05 and p<0.001) the cleavage rate after 30 hours of IVF (53.3%) and at Day 3 (62.7%: Day 0: the day of I VF) compared to control (64.3% and 77.3%, respectively). Subsequent developmental rates to morulae (Mo) and expanding blastocysts (ExBL) also significantly decreased (p<0.001 and p<0.05 for Mo and ExBL) when oocytes were coincubated with sperm in the medium containing MEM amino acids. Taurine added to fertilization medium have not increased the cleavage rate over the control, whereas glycine showed significantly lower (p<0.01) cleavage rate at Day 3 than that of taurine, but there was no significant difference in the developmental rates to Mo and ExBL of bovine embryos irrespective of the supplementation of taurine or glycine to fertilization medium. In conclusion, supplementation of fertilization medium with essential and non-essential amino acids, taurine or glycine has no beneficial effect on in vitro cleavage and development of bovine oocytes matured and fertilization in vitro.

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3-Hyroxyflavone in Maturation Medium Supports In Vitro Development of Fertilized Bovine Follicular Oocytes

  • Kim, Se-Woong;Park, Jong-Im;Jung, Yeon-Gil;Roh, Sangho
    • Reproductive and Developmental Biology
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    • v.38 no.4
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    • pp.143-146
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    • 2014
  • Antioxidants, as reactive oxygen species scavengers, are one of the beneficial additives in serum-free defined culture medium. In this study, three separate experiments were performed to determine the effects of 3-hyroxyflavone added to the culture medium on the developmental competence of follicular bovine oocytes during in vitro maturation (IVM) and/or in vitro culture (IVC). The rate of blastocyst developed from oocytes cultured in IVM medium with 3-hyroxyflavone was significantly higher than that from control oocytes (39.0% vs. 26.3%, p<0.001), respectively. However, oocytes cultured in the medium with addition of 3-hyroxyflavone only at IVC period did not show significance in the blastocyst development when compared with control. When 3-hyroxyflavone was added to both IVM and IVC media, the rate of blastocyst formation was even significantly lower (21.1%) than control (26.5%; p<0.05). The present findings suggested that antioxidative activity of 3-hydroxyflavone added to only IVM medium beneficially affected the developmental competence of follicular bovine.

Studies of In Vitro Embryo Culture of Guppy (Poecilia reticulata)

  • Liu, LiLi;Lee, Ki-Young
    • Development and Reproduction
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    • v.18 no.3
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    • pp.139-143
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    • 2014
  • Different with other fishes, the guppies (Poecilia reticulata) is ovoviviparity, which retain their fertilized eggs within the follicle throughout gestation. The synchronously growing diplotene oocytes store nutrients in droplets and yolk, before their maturation and fertilization. The lecithotrophic strategy of development entails the provisioning of embryos with resources from the maternal yolk deposit rather than from a placenta, it allows the extracorporeal culture of guppy embryo. Studies on their early development of live bearers like the guppy including lineage tracing and genetic manipulations, have been limited. Therefore, to optimize conditions of embryo in vitro culture, explanted embryos from pregnant females were incubated in embryo medium (L-15 medium, supplemented with 5, 10, 15, 20% fetal bovine serum, respectively). We investigated whether the contents of FBS in vitro culture medium impact the development of embryos, and whether they would hatch in vitro. Our study found that in 5% of FBS of the medium, although embryos developed significantly slower in vitro than in the ovary, it was impossible to exactly quantify the developmental delay in culture, due to the obvious spread in developmental stage within each batch of eggs, and embryos can only be maintained until the early-eyed. And although in culture with 20% FBS the embryos can sustain rapid development of early stage, but cannot be cultured for the entire period of their embryonic development and ultimately died. In the medium with 10% and 15% FBS, the embryos seems well developed, even some can continue to grow after follicle ruptures until it can be fed. We also observed that embryonic in these two culture conditions were significantly different in development speed, in 15% it is faster than 10%. But 10% FBS appears to be more optimizing condition than 15% one on development process of embryos and survival rate to larvae stage.

A Comparative Study on the Parthenogenetic Development of Pig Oocytes Cultured in North Carolina State University-23 and Porcine Zygote Medium-3

  • Lee, Joo-Hyeong;Hyun, Sang-Hwan;Lee, Eun-Song
    • Journal of Embryo Transfer
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    • v.27 no.2
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    • pp.121-126
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    • 2012
  • The objective of this study was to examine the effect of in vitro culture media on embryonic development of in vitro-matured (IVM) oocytes after parthenogenetic activation (PA) in pigs. Immature pig oocytes were matured in TCM-199 supplemented with porcine follicular fluid, cysteine, pyruvate, EGF, insulin, and hormones for the first 22 h and then further cultured in hormone-free medium for an additional 22~26 h. IVM oocytes were activated by electric pulses and cultured in porcine zygote medium-3 (PZM-3) and North Carolina State University-23 supplemented with essential and non-essential amino acids (NCSU-23aa). These media were further modified by supplementing 2.77 mM myo-inositol, 0.34 mM trisodium citrate, and $10{\mu}M$ ${\beta}$-mercaptoethanol (designated as mPZM-3 and mNCSU-23aa, respectively). Culture of PA embryos in mPZM-3 significantly increased development to the blastocyst stage than culture in NCSU-23aa (36.2% vs. 24.8%, p<0.05). Modified PZM-3 showed a significantly higher blastocyst formation than NCSU-23aa in both groups of embryos that were activated at 44 h and 48 h of IVM (51.0% vs. 35.5% and 49.0% vs. 34.2% in oocytes activated at 44 h and 48 h of IVM, respectively). Irrespective of the follicle diameter where oocytes were collected, embryonic development to the blastocyst stage was increased (p<0.05) by the culture in mPZM-3 compared to culture in NCSU-23aa (25.9% vs. 34.2% and 32.9% vs. 44.8% in embryos derived from small and medium size follicles, respectively). Our results demonstrated that culture media had significant effect on preimplantation development PA embryos and that mPZM-3 was superior to mNCSU-23 in supporting development to the blastocyst stage in pigs. This beneficial effect of mPZM-3 on embryonic development was not impaired by other factors such as time of oocyte activation and origin of immature oocytes (small and medium size follicles).

A study on learning organization to develop employees' vocational competency at small and medium enterprises and productivity (중소기업 학습조직이 기업 생산성에 미치는 영향에 관한 연구)

  • Lim, Kyung-Sik;Choi, Byung-Gie;Jung, Jang-Woo;Han, Young-Geun;Kang, Kyung-Sik
    • Proceedings of the Safety Management and Science Conference
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    • pp.573-588
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    • 2009
  • In this era of knowledge and information of the 21st century, the source of a nation's competitive power consists of human resources and material resources and the importance of human resources appreciates highly. The number of small and medium enterprises which are the core of national economy covers over 98% among whole domestic enterprises of Korea. Competitive power of small and medium enterprises comes from human resources development. For the development of human resources, small and medium enterprises are trying to expand their learning organizations in the company. The analysis of outcomes of administering the existing learning organizations shows that learning organizations contribute highly to the improvement of the productivity of small and medium enterprises. And the analysis also shows that the participants of learning organizations and middle management and executives who support and encourage them recognize the necessity of learning organizations and that it should be disseminated throughout other enterprises. Therefore government's supports to establish learning organizations in a company are required to expand.

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A study on learning organization to develop employees' vocational competency at small and medium enterprises and improvement of productivity (중소기업 학습조직이 기업 생산성에 미치는 영향에 관한 연구)

  • Lim, Kyung-Sik;Choi, Byung-Gie;Jung, Jang-Woo;Han, Young-Geun;Kang, Kyung-Sik
    • Journal of the Korea Safety Management and Science
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    • v.11 no.3
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    • pp.167-179
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    • 2009
  • In this era of knowledge and information of the 21st century, the source of a nation's competitive power consists of human resources and material resources and the importance of human resources appreciates highly. The number of small and medium enterprises which are the core of national economy covers over 98% among whole domestic enterprises of Korea. Competitive power of small and medium enterprises comes from human resources development. For the development of human resources, small and medium enterprises are trying to expand their learning organizations in the company. The analysis of outcomes of administering the existing learning organizations shows that learning organizations contribute highly to the improvement of the productivity of small and medium enterprises. And the analysis also shows that the participants of learning organizations and middle management and executives who support and encourage them recognize the necessity of learning organizations and that it should be disseminated throughout other enterprises. Therefore government's supports to establish learning organizations in a company are required to expand.

Development of the Enterprise Process Evaluation for Small and Medium Business Consulting (중소기업 경영컨설팅을 위한 전사적 프로세스 평가모델 개발)

  • Cho, Jey-Ryong;Lee, Heejung
    • Journal of the Society of Korea Industrial and Systems Engineering
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    • v.35 no.4
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    • pp.41-47
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    • 2012
  • To adapt to a rapidly changing business environment, introduction of new management techniques and innovative activities are crucial. In the case of small and medium companies, the compensation for scarce management resources and sustainable development can be obtained from advantage of management consulting. In this study, we propose the enterprise process evaluation model for small and medium business consulting, which can deploy the process improvement activities with considering the process maturity level within an organization. The process evaluation area of proposed Small and Medium Business, Staged Maturity Model (SMB-SMM) consists of four following parts: supply management, demand management, business management, and research and development (R&D) management. For each area, we set goals and also define the practices to achieve their goals respectively. By applying the SMB-SMM model to the small and medium sized companies, the process maturity level can be defined. And then to overcome the gap between the goals and evaluation results, the customized management consulting activities can be drawn from the predefined consulting approaches from the SMB-SMM.