• Title/Summary/Keyword: in vitro effect

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Effect of Insulin, Transferrin and Platelet-Derived Growth Factor Supplemented to Synthetic Oviduct Fluid Medium on In Vitro Development of Bovine Embryos Matured and Fertilized In Vitro (합성난관배양액에 첨가된 Insulin, Transferrin 및 Platelet-Derived Growth Factor (PDGF)가 소 수정란의 체외발육에 미치는 영향)

  • 이은송
    • Journal of Embryo Transfer
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    • v.12 no.3
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    • pp.283-291
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    • 1997
  • In vitro development of bovine embryos is affected by many factors such as energy substrates, amino acids, and some growth factors. It has been reported that mRNA of insulin, PDGF and their receptors are detected in cow embryos, and that some chelating agents such as EDTA and transferrin have beneficial role on mouse and bovine embryos. The author hypothesized that insulin, transferrin arid PDGF added to a culture medium increase in vitro development of bovine embryos by chelating toxic substance(s) or increasing cell growth and metabolism. Immature oocytes from slaughtered ovaries of Holstein cows and heifers were matured for 24 hours in a TCM199 containing 10% fetal calf serum, FSH, LH and estradiol with granulosa cells in vitro. Matured oocytes were coincubated with sperm for 30 hours in a modified Tyrode's medium (IVF). Embryos cleaved to 2- to 4-cell at 30 hours after IVF were selected and cultured in a 30-$\mu$l drop of a synthetic oviduct fluid medium (SOFM) containing 0.8% BSA, Minimum Essential Medium essential and non-essential amino acids, and insulin, transferrin or PDGF for 9 days. Supplementation of a SOFM with insulin, and /or transferrin did not increase develop-mental rate to expanding and hatching blastocyst of 2- to 4-cell bovine embryos compared with control. The highest developmental rate to hatching blastocyst was shown when PDGF was added at the concentration of 10 ng /ml among the supplementing doses tested in the present study (p<0.05). Addition of PDGF without insulin to a SOFM could not increase embrye development, but combined addition of PDGF with insulin significantly increased (p<0.05) embryo development to hatching blastocyst (50%) compared with control (38%). In conclusion, insulin and PDGF supplemented to a SOFM may act synergistically and have beneficial effect on in vitro development of 2- to 4-cell bovine embryos matured and fertilized in vitro.

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Cytocidal Effect of Hyperthermia on Tumor Cells in vivo (In vivo 腫瘍細胞에 미치는 溫熱處理의 細胞致死效果)

  • Kang, Man-Sik;Rhee, Jeong-Gile;Seymour H. Levitt;Chang W. Song
    • The Korean Journal of Zoology
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    • v.24 no.2
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    • pp.59-64
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    • 1981
  • The cytocidal effect of hyperthermia on subcutaneous SCK tumor cells growing in vivo was significantly greater than that on the SCK tumor cells cultured in vitro. When the tumors were left in situ after heating, the cell survival progressively decreased, and the functional intratumor vascular volume also decreased. The radiation survival curves of tumor cells heated either 30 min before or after X-irradiation in vivo were steeper than the radiation survival curves of unheated control tumors. It is concluded that the cytocidal effect of hyperthermia on tumor cells in vivo is greater than that in vitro due possibly to the intratumor environment.

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The Effect of Polyamines on the DNase Activity in Cultured Carrot Cells (당근(Daucus carota L.)배양세포의 DNase활성에 미치는 Polyamines의 영향)

  • 윤미정
    • Journal of Plant Biology
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    • v.29 no.4
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    • pp.285-294
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    • 1986
  • The present study was attempted to investigate the effects of polyamines such as putrescine, spermidine and spermine on protein content and DNase activity in vivo and in vitro in carrot embryos. It was also investigated whether polyamines could replace role of cations required for DNase activity in vitro. The results obtained are as follows. Putrescine, spermidine and spermine increased protein content, although response to spermine reached plateau at the concentration of 0.1 mM. DNase activity was inhibited by polyamines, the inhibition being concentration-dependent and the highest att he concentration of 10 mM. The inhibition of DNase activity was the most prominent with spermine. Similar inhibitory effect to polyamines which was concentration-dependent was found in DNase activity but no change was shown on time-course in vitro. Putrescine and spermidine enhanced the DNase activity at low Mg2+ and Mn2+ concentrations, suggesting that the role of Mg2+ and Mn2+ for DNase activity could be, in part, replaced by these polyamines. These results, therefore, suggest that plyamines can modulate DNase activity through binding to DNA rather than direct effect on DNase activity.

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Effects of Insulin, Transferrin and Selenium (ITS) on In Vitro Development of Porcine Parthenogenetic and Nuclear Transfer Embryos

  • Quan, Yan-Shi;Naruse, Kenji;Kim, Baek-Chul;Kim, Hong-Rye;Han, Rang-Xun;Choi, Su-Min;Park, Chang-Sik;Jin, Dong-Il
    • Reproductive and Developmental Biology
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    • v.31 no.4
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    • pp.261-265
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    • 2007
  • Insulin, transferrin and selenium (ITS) complex is reported to improve in vitro development of oocytes and embryos. This study was carried out to investigate the effects of ITS during in vitro culture (IVC) of porcine parthenogenetic and nuclear transfer (NT) embryos on subsequent developmental capacity in vitro. The electrically activated oocytes were cultured in Porcine Zygote Medium (PZM-3) with various concentrations (0, 0.1, 0.5, and 1.0%) of ITS for 7 days. Also, the electrically activated reconstructed embryos were cultured in PZM-3 with various concentrations (0, 0.1, 0.5, and 1.0%) of ITS for 6 days. Addition of ITS to culture medium did not affect development of porcine parthenogenetic embryos in vitro. To test the effect of ITS on the in vitro development of porcine NT embryos, factorial experiments were also performed for in vitro maturation (IVM) medium (TCM-199) with or without 1% ITS and culture medium (PZM-3) with or without 0.5% ITS. Addition of 0.5% ITS to culture medium increased (p<0.05) the proportion of NT blastocysts compared with non-treated group. In contrast, addition of 1% ITS to culture medium was ineffective or had a detrimental effect. Also, addition of ITS only to maturation medium increased (p<0.05) the percentage of NT blastocysts formation compared with the control group. In conclusion, addition of ITS to IVM or IVC medium could improve subsequent blastocyst development of porcine NT embryos.

Effect of Puerariae Radix Methanol Extract on Benzo(a)pyrenc -in - duced Hepatotoxicity in Rats (갈근 메탄올 엑기스가 흰쥐에 있어서 Benzo(a)pyrene에 의해 유도된 간장해에 미치 는 영향)

  • 이윤경
    • Journal of the East Asian Society of Dietary Life
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    • v.4 no.2
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    • pp.59-67
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    • 1994
  • The present study was conducted to evaluate the hepatoprotective effect of puerariae Radix methanol extract on benzo(a) pyrene(B(a)P) - induced liver injuries in rats. In vitro experiment, primary cultured hepatocytes (5X105 cells/$m\ell$) were cultured for 20~24 hours after adding puerariae Radix mehtanol extract(32$\mu\textrm{g}$/$m\ell$) and B(a)P(50 uM). In vivo experiment, Puerariae Radix methanol extract(0.25 g/kg/day, per os) was administered for 7 days and B(a)P(0.1 mg/kg/day, intraperitoneally) was given after the last administration of extract. And then the hepatoprotective effect of Puerariae Radix methanol extract was investigated biochemically through in vitro and in vivo experiments. Namely, activities of enzymes (GOT, GPT and LDH) were measured and 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide(MTT) assay were carried out in vitro cell culture study and GOT, GPT, LDH and ALP activities and HDL-cholesterol, total cholesterol and triglyceride contents were performed in vivo study. In vitro experiment, as a result of enzyme activity measurement(GOT, GPT and LDH) and MTT assay, GOT,GPT and LDH activities changed by B(a)P were recovered to normal levels and hepatocytes impaired by B(a)P were recovered to normal. In vivo experiment, Puerariae Radix methanol extract significantly decreased the enzyme activities(GOT, GPT, ALP and LDH in serum and GPT and ALP in tissue) and lipid contents in comparison to B(a)P-treated group.

In vitro Screening of UVA Phototoxicity Inhibitors using the Natural Products (In vitro 실험법에 의한 천연물 중의 UVA 광독성 억제제 검색)

  • 김현진;김봉희
    • Environmental Analysis Health and Toxicology
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    • v.17 no.3
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    • pp.253-259
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    • 2002
  • The phototoxicity inhibitory activity of 15 natural products having antiinflammatory effect was screened by three in vitro methods: yeast growth inhibition test with Candida albicans, RBC photohemolysis and MTT assay. We induced phototoxic reaction by irradiating UVA (365 nm) on chlorpromazine (CPZ) that has been widely documented as phototoxic agent in clinical and experimental studies and then observed the effects of the natural products after treating them with CPZ. In yeast growth inhibition test, X. stramonium showed the inhibitory effect on the UVA phototoxicity and E. officinalis, Yeast, P. suffruticosa showed phototoxicity inhibitory effect in that their % hemolysis compared with control were 36.14${\pm}$ 2.69, 42.82${\pm}$1.35, 36.41${\pm}$0.48 on UVA. In MTT assay, all tested natural products increased cell viability compared with the control.

Proapoptotic and antitumor effect of Hangbaek-Tang(HBT) in a tumor transplanted mouse model (마우스 모델에서 항백탕 투여에 의한 종양 증식의 억제 및 Apoptosis의 유도)

  • Yun, Young-Gab;Kim, Jun-Hee;Song, Eun-Jung;Hwang, Jin-Ki;Nam, Sang-Yun
    • Herbal Formula Science
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    • v.17 no.2
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    • pp.73-83
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    • 2009
  • Objective : In vitro proapoptotic effect of Hangbaek-Tang (HBT) has been documented by one of us. In the present study, we aimed to demonstrate in vivo effect of HBT on tumor growth. Methods : In vitro selective cytotoxicity of HBT was examined by enumeration of viable cell numbers using BC3A mouse leukemic cells and normal spleen cells. In vivo effect of HBT (25 and 50 mg/mouse) on tumor growth was assayed using BC3A cells innoculated subcutaneously in the flank. Annexin-V apoptosis assay and PI staining was performed to determine the effective serum factor in HBT-treated mice. Leukocyte recruitment into peritoneum were analyzed by microscopy with a stained cytosmear of peritoneal lavage fluid. Results : HBT exhibited in vitro selective cytotoxicity to leukemic cells and did not show any toxicity on immune organs. In vivo i.p. administration of HBT induced significant reduction in tumor growth but not complete regression. Sera obtained from HBT-treated mice strongly inhibited BC3A cell growth in vitro and were revealed to markedly enhance apoptosis and accompanying cell death, when compared to those from PBS-treated mice. Abundant extravasation of leukocytes, especially neutrophils, into peritoneum was observed in HBT-treated mice. Conclusions : HBT causes leukemic, BC3A cell death in vivo via apoptosis as well as in vitro, for which functional involvement of leukocytes is suggested.

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Effect of Antioxidants on In Vitro Development of Korean Native Cattle Embryos Derived from In Vitro Fertilization (항산화제 첨가가 한우 체외 수정란의 체외 배발달에 미치는 영향)

  • 문승주;김은국;김재홍;명규호;선상수
    • Journal of Embryo Transfer
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    • v.14 no.3
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    • pp.219-224
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    • 1999
  • The effect of several potential antioxidants were examined as a means of increasing the in vitro development of in vitro matured and in vitro fertilized oocytes into morulae and blastocysts. Korean native cattle embryos after in vitro fertilization were cultrued for 7 days at 38.5$^{\circ}C$ in CR1aa containing varing concentration of the antioxidants in a gas phases consisting of 5% CO2, 95% humidified air. The results obtained were summarized as follows; The proportion of embryos developed to morulae and blastocysts in CR1aa containing 2.5uM $\alpha$-tocopherol(11.0% and 6.0%) was significantly higher than those of 0, 5.0, and 7.5uM $\alpha$-tocopherol (P<0.05). concentration of 50uM L-ascorbic acid (7.5% blastocysts) did affect the proportion of embryos developing into blastocystes(P>0.05). Addition of 200uM cysteamine was significantly higher than those of 0, 100 and 300uM (P<0.05). When the fertilized oocytes were cultured at 0. 200, 400 and 600uM of selenium for 168 hrs, the morulae rates were 12.2, 5.2, 16.0 and 16.1% respectively, and addition of 200uM selenium was significantly higher than those of 0, 400, 600uM (P<0.05). These results suggested that the addition of $\alpha$-tocopherol, L-ascorbic acid, cysteamine and selenicum can enhanced development to the morulae and blastocysts of in vitro derived fertilized oocytes.

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In vitro cell recovery methoc as an altermative to human damaged skin recovery test

  • An, Su-Sun;Nam, Ki-Taek;Park, Jong-Ho;Koh, Jae-Sook
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.23 no.3
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    • pp.97-100
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    • 1997
  • These days, the raw materials that have the cell recovering effect are used commonly in cosmetics. In this study, six materials were rested for the characteristics of recovering effect both on vivo and in vitro. Tested raw materials were Soypol, 3-APPA, Apple extract, Polygonatum japonicum extract, Scutellarkd baicalensis extract, Aloe extract. Among these materials, Soypol and 3-APPA were synthesized and others were made by extraction at the Pacific R&D Center. Human forearm skin and cultured skin cell were damaged by sodium lauryl sulfare and then raw materials were applied for open treatment on SLS damaged human skin or cells. The recovering effects of raw materials in vivo were evaluated by measuring transepidermal water loss, skin hydration and erythema and in vitro effects of proliferationg cells were assessed by neutral red uptake assay. In the in vivo study, only the evaluation by TEWL showed correlation with the visual score. Our of six materials, 3-APPA had the most positive effect in both in vivo and in vitro studies and the correlation was r=0.8286 (p=0.042).

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Epidermal Growth Factor Induces Bcl-xL Gene Expression and Reduces Apoptosis in Porcine Diploid Parthenotes Developing in vitro

  • X. S. Cui;M. R. Shin;S. H. Jun;Kim, N. H.
    • Proceedings of the KSAR Conference
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    • 2003.06a
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    • pp.53-53
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    • 2003
  • The aim of this study was to determine the interactive effects of BSA and EGF on the viability and development of porcine diploid parthenotes developing in vitro. The addition of 0.1 and 0.4% BSA to the culture medium enhanced the development of 4-cell parthenotes to the blastocyst stage but EGF had no effect. However, while BSA also increased cell numbers, it did so only when EGF was also present. Either agent on its own had no effect. Similarly, apoptosis in the blastocysts was not influenced by either agent on its own but was reduced when both BSA and EGF were present. Furthermore, semi-quantitative reverse-transcriptase polymerase chain reaction (RT-PCR) revealed that EGF enhanced the mRNA expression of BclxL in the presence of 0.4% BSA but BSA and EGF alone had no effect. EGF and/or BSA did not influence Bak gene expression in the blastocyst stage parthenotes. These results suggest that BSA has both beneficial and detrimental effects on the viability of porcine diploid parthenotes developing in vitro and that exogenous EGF may block some of the detrimental effects of BSA, possibly by inhibiting the BSA-induced apoptosis by increasing Bcl-xL expression. This results in a net increase in cell numbers in porcine diploid parthenotes developing in vitro.

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