• Title/Summary/Keyword: cultured media

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The Effect of Nitric Oxide on the Embryonal Development in Mouse (NO(Nitric Oxide)가 생쥐의 배 발달에 미치는 영향)

  • Min, Bu-Kie;Kim, Kie-Suk;Rhee, Hee-Sub;Hong, Gi-Youn;Shin, Hyeong-Do;Sung, Yeon-Kyeong;Kim, Hyung-Min
    • Clinical and Experimental Reproductive Medicine
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    • v.25 no.2
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    • pp.109-113
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    • 1998
  • Objective: To ananlyze the direct effect of nitric oxide (NO), generated from sodium prusside (SNP) on the embryo developments in reproductive process. Design: Ova from mouse were treated to allow fertilization in in vitro culture. And the samples of fertilized ova were alloted into five alliqutos. Each alliquot was cultured in media treated with either concentration at 0 (n=92), $25{\mu}M$ (n=84), $50{\mu}M$ (n=80), $100{\mu}M$ (n=77), $500{\mu}M$ (n=54) of SNP. Main Outcome Measure: Rates of embryonal cell cleavages, viability and cell morphology were assessed during in vitro fertilization and culture. Results: As analyse the cell cleavage at 24 hours after in vitro culture of fertilised egg in variuos NO concentration, all of egg cells of each alliquot were developed to $2\sim4$ cell stage. But the alliquot of egg cells treated with $50{\mu}M$, which were totally degenerated. And also all embryonal cells of each alliquot were developed to 8 cell stage and morula stage on culture continuosly. And the embryonal cells of each alliquot were analysed at 24 and 48 hours following the in vitro culture. The rates of cell fragmentation and fusion were $4.2{\pm}3.4%$ in control group which is not treated with NO, while experimental groups was high, as rated $23.4{\pm}6.2%$ in $25{\mu}M$, $28.2{\pm}5.7%$ in $50{\mu}M$ and $32.1{\pm}6.4%$ in $100{\mu}M$ concentration of NO. Accordingly the rate of abnormal morphology of embryonal cell in control was lower significantly than that in each alliquot of experimental groups (p<0.05). And the degenerated rates of embryonal cells were 0% in control, $17.8{\pm}6.7%$ in $25{\mu}M$, $23.6{\pm}4.7%$ in $50{\mu}M$ and $26.8{\pm}11.2%$ in $100{\mu}M$ at 8 cells and morula on culture of 48 and 72 hours. On the examination of embryonal cells developed to blastocyst through in vitro culture, the rates of degenerated cells were $16.8{\pm}7.2%$ in control, $37.5{\pm}6.2%$ in $25{\mu}M$, $73.4{\pm}4.6%$ in $50{\mu}M$, 100% in $100{\mu}M$. Conclusion: This results suggeted that the NO in any concentrations is harmful on embryos in view of morphology as well as viability of cell, and the toxicity of NO on embryo is stronger at condition in higher concentration of NO.

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Effects of Medium on Blastocyst Formation, Cell Number and Percentage of ICM in Mice (마우스에서 배반포 형성, 세포 수 및 ICM의 비율에 미치는 배양액의 효과)

  • Park, Kee-Sang;Lee, Taek-Hoo;Chun, Sang-Sik;Song, Hai-Bum
    • Clinical and Experimental Reproductive Medicine
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    • v.29 no.4
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    • pp.237-243
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    • 2002
  • Objective : The aim of this study was to evaluate the influence of different media on blastulation, mean cell number, percentage of inner cell mass (ICM) of total cells and ICM : trophectoderm (TE) ratio in mice. Materials and methods: A total 552 two cell embryos were retrieved from ICR female mice (4 weeks old) at 48 hr after hCG injection (mated just after hCG injection) and cultured in MEM (n=276) or TCM (n=276) supplemented with 20% hFF. The grading of blastocysts from zona-intact (ZiB) to -escape (hatching and hatched, ZeB) was performed at 72 hours after culture. Total, TE and ICM cell numbers were analyzed by differential staining of blastocyst. Statistical analysis was performed using t-test with SigmaPlot-2001. P-values < 0.05 were accepted as statistically significant. Results: The blastulation rate in MEM ($64.9{\pm}4.95%$) was significantly higher (p=0.0031) than that in TCM ($57.2{\pm}5.22%$). No differences were found in the number of ZiB and ZeB between MEM ($31.9{\pm}2.62$, $33.0{\pm}4.58%$), and TCM ($27.2{\pm}4.28$, $30.1{\pm}4.58%$). A total 314 blastocysts (MEM=166; TCM=148) were stained differentially. Mean cell number of blastocysts was significantly higher (p=0.0002) in TCM ($73.1{\pm}3.3$) than in MEM ($61.7{\pm}2.5$). Differential staining was successfully performed in 155 blastocysts (MEM=77; TCM=78). The percentage of ICM was significantly higher in MEM than in TCM ($20.9{\pm}1.3$ vs. $17.1{\pm}1.2%$, p=0.0281). The ICM : TE ratio was higher in TCM than in MEM (1 : $4.85{\pm}0.68$ vs. 1 : $3.78{\pm}0.78$, NS). Conclusion: These results show that MEM increase the blastocyst formation and percentage of ICM of total cells comparing with TCM in mice.

Comparison of the clinical outcomes of day 4 and 5 embryo transfer cycles

  • Lee, Sun-Hee;Lee, Hyoung-Song;Lim, Chun Kyu;Park, Yong-Seog;Yang, Kwang Moon;Park, Dong Wook
    • Clinical and Experimental Reproductive Medicine
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    • v.40 no.3
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    • pp.122-125
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    • 2013
  • Objective: The majority of embryo transfers (ETs) to date have been performed on day 3 to reduce the potential risk of developmental arrest of in vitro cultured embryos before ET. Development of sequential media has significantly improved culture conditions and allowed blastocyst transfer on day 5. While day 5 ET provides higher clinical pregnancy outcomes with reduced risks of multiple pregnancies, it still has potential risks of developmental arrest of IVF embryos. The aim of this study was to evaluate the clinical outcomes of day 4 ETs and compare the efficacy of day 4 ET with day 5 ET. Methods: From 2006 to 2009, a total of 747 fresh IVF-ET cycles were retrospectively analyzed (day 4, n=440 or and day 5, n=307). The cycles with any genetic factors were excluded. The rates of matured oocytes, fertilization, good embryos, and clinical pregnancy of the two groups were compared. The chi-square test and t-test were used for statistical analysis. Results: There were no significant differences between the two groups with respect to the mean age of the females and rates of matured oocytes. The pregnancy outcomes of day 4 ET (40.7%) were similar to those of day 5 ET (44.6%). The implantation rate of day 5 ET (24.2%) was significantly higher than that of day 4 ET (18.4%) (p=0.003). Conclusion: Day 4 ET can be chosen to avoid ET cancellation in day 5 ET resulting from suboptimal circumstances in the IVF laboratory, but the decremented quality of embryos for transfer and the decreased pregnancy rate must be taken into consideration.

Anti-bacterial and Anti-fungal Effects of Herbal Oil Made from Vateria acuminata Hyne (Vateria acuminata Hyne으로 만든 오일의 항세균 및 항진균 효과)

  • kim, Soo-Ji;Cabral, L.M Udaya;Hong, Jin-Young;Jo, Chang-Wook;Kim, Young-Hee;Choi, Jung-Eun
    • 보존과학연구
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    • s.33
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    • pp.5-17
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    • 2012
  • An Ola leaf manuscript, which consists of words carved on an Ola leaf and is filled up with a mixture of herbal oils made from Vateria acuminata Hyne and charcoal, is one of the native writing media in Sri Lanka. According to historical records, Ola leaf manuscripts had been used from the 1st to the 18th century A.D. From the recent findings that Ola leaf manuscripts have been preserved well for the past 400 years and are in good state of preservation, it is supposed that herbal oils preserve Ola leaves against environmental and biological factors such as fungi and insects. To evaluate the anti-biological susceptibility of the herbal oils, the molds isolated from wooden printing blocks in Janggyeong Panjeon of Haeinsa Temple in South Korea and bacteria and fungi isolated from Ola leaves were cultured. After spreading the microorganisms suspension on an agar plate, a disk paper containing a certain volume of herbal oil was placed on the agar plate. It was found in the experiment that herbal oil exhibits a clear zone, which is optically clear and inhibits the growth of microorganisms, against some molds and bacteria. The study results indicate that the herbal oil from the plant Vateria acuminata Hyne has the anti-bacterial and anti-fungal properties.

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Interaction of Schwann Cells with Various Protein- or Polypeptide-Coated PLGA Surfaces (다양한 단백질과 폴리펩타이드로 코팅된 PLGA 표면과 슈반세포와의 상호관계)

  • Park Ki-Suk;Kim Su-Mi;Kim Moon-Suk;Lee Il-Woo;Rhee John-M.;Lee Hai-Bang;Khang Gil-Son
    • Polymer(Korea)
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    • v.30 no.5
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    • pp.445-452
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    • 2006
  • In this study, we investigated interaction of Schwann cells (SCs) with various cell-adhesive coated polymer surface. We used cell-adhesives that like a fibronectin (FN), fibrinogen(FG), laminin(LM), vitronectin (VN), poly-D-Iysine (PDL), and poly-L-Iysine (PLL) to coat PLGA film surface and evaluated the surface property of coated or not PLGA films by measurement of water contact angle and ESCA. SCs were cultured on coated or non-coated PLGA film surface, and then examined the cell adhesion and proliferation by cell count and SEM observation. Cell count results revealed initial cell adhesion related to protein adsorption on PLGA surface. In addition, serum content in media related to cell proliferation rate. In this result, we recognized that adhesion and proliferation of SCs were affected by specific cell-adhesives. In these results, we recognized that is important to provide the suitable surface environment according to cell types and culture condition for improvement of cell adhesion and proliferation.

Effect of Plant Growth Regulators and Media on Regeneration of Sorghum bicolor (L.) Moench (바이오에너지용 수수 품종의 재분화율 증진을 위한 배지와 생장조절제 효과)

  • Goh, Eun-Jeong;Seong, Eun-Soo;Yoo, Ji-Hye;Kil, Hyun-Young;Lee, Jae-Geun;Hwang, In-Seong;Kim, Nam-Jun;Ghimire, Bimal Kumar;Kim, Myong-Jo;Lee, Ju-Kyung;Lim, Jung-Dae;Kim, Na-Young;Yu, Chang-Yeon
    • Korean Journal of Plant Resources
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    • v.24 no.2
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    • pp.168-173
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    • 2011
  • This study was carried out to optimize the embryogenic callus induction and plant regeneration from mature seeds of Sorghum bicolor. The effect of growth regulators was investigated on formation of embryogenic callus. The highest frequency of embryogenic callus was observed when the mature seeds were cultured on B5 medium supplemented with 2 mg/L 2,4-Dichlorophenoxyacetic acid (2,4-D). The highest frequency of plant regeneration from embryogenic callus was observed on MS medium with 0.5 $mg\;l^{-1}$ 6-benzyl amino purine (BAP) and 0.25 $mg\;l^{-1}$ indole-3-butyric acid (IBA) to optimize the shoot regeneration. High concentration of BAP (1 $mg\;l^{-1}$) supplemented with IBA (0.25 $mg\;l^{-1}$) was effective combination for shoot multiplication. MS medium supplemented with 1 $mg\;l^{-1}$ IBA was found to be the most effective for inducing roots. Normal rooted plantlets were transferred to the greenhouse for hardening with over 90% survival rate. Hence, this reproducible protocol could be useful for mass propagation and genetic transformation of S. bicolor.

The Effects of Cyclic AMP, Theophylline and Caffeine on Sporulation of Aspergillus niger in the Defined Media (검정곰팡이(Aspergillus niger)의 포자형성(胞子形成)에 미치는 cAMP, 테오필린 및 카페인의 영향(影響)에 관한 연구(硏究))

  • Koh, Young-Joo;Kim, Jong-Hyup
    • The Korean Journal of Mycology
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    • v.15 no.4
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    • pp.238-246
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    • 1987
  • Aspergillus niger van Tieghem was cultured by the method of synchronous and submerged culture. The sporulation occurred through the culture and its life cycle and differentiation were completed in the experiments. The effects of cAMP, theophylline and caffeine on the sporulation of A. niger were investigated. In the sporulation medium, the sporulation was stimulated by addition of cAMP and its optimum concentration was $10^{-4}M$. In the sporulation medium, the sporulation was stimulated by addition of theophylline and its optimum concentration was 10 mg/ml. In the sporulation medium, the sporulation was stimulated by addition of caffeine and its optimum concentration was 300 mg/ml. Theophylline added to the sporulation medium together with cAMP enhanced the promotion effect of cAMP on sporulation. Caffeine added to the sporulation medium together with cAMP enhanced the promotion effect of cAMP on sporulation. In the sporulation medium, the sporulation was stimulated by addition of neither AMP nor ATP. In the potassium acetate medium, cAMP, theophylline and caffeine stimulated the sporulation, respectively.

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The Condition of Production of Artificial Fruiting Body of Cordyceps militaris (Cordyceps militaris 인공자실체 형성조건)

  • Choi, In-Young;Choi, Joung-Sik;Lee, Wang-Hyu;Yu, Young-Jin;Joung, Gi-Tae;Ju, In-Ok;Choi, Young-Keun
    • The Korean Journal of Mycology
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    • v.27 no.4 s.91
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    • pp.243-248
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    • 1999
  • To compare the morphological and cultural characteristics of Cordyceps militaris, forming conditions of artificial fruiting body production were investigated in this experiment. The stroma shapes of artificial fruiting body were confirmed to club and/or stick. However, both shapes were same color as a orange. Perithecia of stroma was semi-egg shape and was ranged from $30{\sim}90{\times}90{\sim}130\;{\mu}m$ and it's asci was narrowly cylindric. Ascospore of perithecia was belong to filiform and multiseptate. The conidia of C. militaris was centurally grown and shaped with globose, long clavate type, floccose, centrally raised during anamorph stage. When it was cultured between glass and PP bottle, production of artificial fruiting body, pinheading ratio and total yield were higher in PP bottle. The optimum illumination was 1,000 lx for the those production. The culture medium of barley, wheat and hulled rice showed higher artificial fruiting body compared to that of silkworm. Pinheading and yield of it's isolates was decreased at more than three subsequent transculture.

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Antioxidant Activity and Inhibitory Activities of Xanthine Oxidase and Tyrosinase of Yeasts from Wild Flowers in Korea (우리나라 야생화에서 분리한 효모의 항산화 활성과 Xanthine Oxidase 및 Tyrosinase 저해활성)

  • Han, Sang-Min;Hyun, Se-Hee;Kim, Na-Mi;Lee, Jong-Soo
    • The Korean Journal of Mycology
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    • v.43 no.2
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    • pp.99-103
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    • 2015
  • We prepared supernatants and cell-free extracts of yeasts isolated from wild flowers in Korea, and their antioxidant activity and inhibitory activities of xanthine oxidase and tyrosinase were investigated. Among them, cell-free extracts of Kuraishia capsulata UL40-2 and Sporobolomyces ruberrimus 121-Z-3 showed significantly high xanthine oxidase inhibitory activity of 46.4% and 48.3%, respectively. Starmerella bombicola 80-J-1 also showed tyrosinase inhibitory activity of 36.2% in the cell-free extract. Other antioxidant activities and tyrosinase inhibitory activities were not detected or were below 20%. Maximal production of the xanthine oxidase inhibitors were observed when Kuraishia capsulate UL40-2 and Sporobolomyces ruberrimus 121-Z-3 were cultured in the yeast extract-peptone-dextrose media at $30^{\circ}C$ for 24 hour, respectively.

The Inhibitory Effects of Intestine-oriented Lactobacillus sp. KP-3 on the Accumulation of Heavy Metals in Sprague Dawley rats (Sprague Dawley 쥐에서 장내 유래 Lactobacillus sp. KP-3의 중금속 축적 저해 효과)

  • Kim, Shin Yeon;Kim, Hyun Pyo
    • Journal of Life Science
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    • v.25 no.2
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    • pp.164-173
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    • 2015
  • To investigate the effect of lactic acid bacteria on the heavy metal adsorption from internal organs and blood, lactic acid bacteria were isolated from human feces. Some strains resistant to heavy metals were selected by incubation in agar media containing each of chrome and cadmium salts. Among them, a strain named KP-3 was ultimately chosen due to its higher growth rate in selective broth medium containing the heavy metals at the concentration of 0.01%. The strain was identified as Lactobacillus sp. based on its morphological, cultural and physiological characteristics. For evaluating the ability to prevent accumulation of heavy metals by selected Lactobacillus sp. strain in vivo, Sprague Dawley rats were fed with heavy metal salts (cadmium, chrome and lead) with or without cultured whole cells for 7 days. The amounts of heavy metals accumulated in liver, kidney and blood were analyzed. As a result, chrome was accumulated to kidney mostly, and lead was frequently found in liver and kidney. Experimental group (rats fed with lactic acid bacteria) showed less accumulation of heavy metal than control group (rats fed with saline solution). The inhibition rates of heavy metal accumulation were calculated to 41.8% (Cd), 33.4% (Cr) and 44.2% (Pb). Especially, feeding lactic acid bacteria strongly reduced accumulation of cadmium in blood. The results showed that feeding Lactobacillus sp. KP-3 could prevent the bioaccumulation of heavy metals in the living body.