• Title/Summary/Keyword: in vitro effect

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Effect of the Presence of Corpus Luteum on Oocyte Recovery and Subsequent in vitro Maturation and Fertilization in Buffaloes

  • Singh, Sajjan;Dhanda, O.P.;Malik, R.K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.14 no.12
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    • pp.1675-1677
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    • 2001
  • The effect of the presence or absence of corpus luteum in the ovaries of slaughtered buffaloes was studied for the oocytes recovery and their subsequent maturation and fertilization in vitro. On an average, 0.41 and 0.67 oocytes per ovary were recovered from ovaries with and without corpus luteum, respectively. Immature oocytes were matured in TCM-199 medium. Significant difference was observed in maturation rate between good (74%) and fair (37%) oocytes. However, there was no significant difference in cleavage rate between the two types. The results of this study show that although the presence of corpus luteum in the ovary at the time of recovery significantly affected availability of total oocytes and in-vitro maturation, but fertilization and cleavage remained unaffected under in vitro conditions.

Effect of Bovine Oviductal Epithelial Cell(BOEC) Conditioned Medium on In Vitro Development of Bovine Embryos Fertilized In Vitro (난관상피세포 Conditioned Medium이 체외수정된 소 수정란의 체외 발달에 미치는 영향)

  • 오종훈;김동훈;정형민;이훈택;정길생
    • Korean Journal of Animal Reproduction
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    • v.17 no.1
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    • pp.69-74
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    • 1993
  • This study was investigated to examine the effect of conditioned medium from bovine oviductal cell(BOEC) in the co-culture system with BOEC on in vitro development of in vitro produced bovine embryos. Oocyte-cumulus complexes were cultured for 24 hrs in TCM-199 supplemented with 10% fetal calf serum, 1$\mu\textrm{g}$/ml FSH and 21U hCG, 1$\mu\textrm{g}$/ml oestradiol-17$\beta$ at 39$^{\circ}C$ under 5% CO2 in air. In vitro fertilization was performed with epididymal sperm and heparin (10$\mu\textrm{g}$/ml, 15min.) or caffeine(2.5mM)-treated spermatozoa. Oocytes were incubated with 1$\times$106 spermatozoa/ml for 18 hrs and then cultured in various culture system for 7 days. The development rates of 16-cell or blastocyst stages were recorded on 4, 7 days, respectively, after incubating. The proportions ofembryonic development into molulae and blastocysts were higher in cumulus cell co-culture(23.4%) and BOEC co-culture(34.3%) than in M199-FCS(6.1%). Similarily, the development rates into molulae and blastocysts were significantly higher in BOEC-conditioned medium than those in M199-FCS. Therefore, it is suggested that BOEC co-culture and BOEC conditioned medium increase significantly the development of in vitro produced bovine embryos in in vitro system.

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Effect of Follicle Size in Jeju Mare on Oocytes Matured in vitro (제주말에서 난포 크기에 따른 난포란의 체외성숙)

  • Yoo, Jae-Gyu;Kang, Tae-Young
    • Journal of Veterinary Clinics
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    • v.27 no.1
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    • pp.46-49
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    • 2010
  • In this study, we investigated the number of follicles, oocyte recovery rate and oocyte competence after in vitro maturation according to the size of follicle. And equine oocyte competence after in vitro maturation was investigated in terms of the diameter of follicle with criteria of maturation: nuclear stage after Hoechst staining. The average number of follicles per ovary with middle size (11-20 mm, 2.68) was higher than those of small (5-10 mm, 0.74) and large size follicle (> 21 mm, 1.63), therefore medium follicle (53.1%) had higher proportion than other size of follicles. The average numbers of follicle per ovary was 5.05. The rate of oocyte recovery in small (54.5%) and middle follicle (50%) was higher than that in large follicle (40.9%). After culture for 48 h in Medium 199, 50%, 45.5%, and 44.4% of oocytes from the follicles with diameters of 5-10, 11-20, > 21 mm, respectively reached the metaphase II stage. This is the first report showing number of follicle, oocyte recovery rate according to follicular size, and in vitro oocyte maturation in Jeju mare in Korea. To fulfill in vitro equine embryo production, further studies such as the seasonal effect, in vitro fertilization etc is need.

In Vitro Glucose and Bile Acid Retardation Effect of Fucoidan from Laminaria japonica (다시마 유래 Fucoidan의 In-vitro 포도당 및 담즙산 흡수지연 효과)

  • Park, Kap-Yong;Back, Jin-Hong;Hur, Won;Lee, Shin-Young
    • KSBB Journal
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    • v.22 no.4
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    • pp.265-269
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    • 2007
  • Fucoidan from sea tangle (Laminaria japonica) was isolated by hot water extraction, and partially purified. The in-vitro glucose and bile acid retarding effects of the partially purified fucoidan were investigated. Fucoidan exhibited 27.06$\sim$21.42% of retarding index for glucose and 33.50$\sim$27.02% of retarding index for bile acid during in vitro dialysis experiment for 2 hours. These retarding effects on glucose and bile acid diffusion was considered as a relatively good or very good, suggesting the prevention from diabetes and arteriosclerosis of some extent.

The Effect of Serum and Tramsferrin on the Growth of Malassezia pachydermatis in vitro (In vitro에서의 Malassezia pachydermatis의 성장에 대한 혈청과 Transferrin의 억제효과)

  • 김진영
    • Journal of Veterinary Clinics
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    • v.19 no.1
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    • pp.86-91
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    • 2002
  • The inhibitory effect of pooled canine serum on the growth of ten strains of Malassezia pachydermatis in vitro was investigated. Studies were also carried out to observe the effect of different concentrations of unsaturated bovine transferrin on Malassezia growth in vitro. Ten strains of Malassezia pachygedrmatis in normal canine serum (11.1%, 44.4%) were found to significantly inhibit the growth (p<0.0005) not only in a dose dependent but also in a time dependent manner. The same strains of yeast treated with 1, 2, 4 and 8 times the normal value of serum transferrin (3.0 mg/ml, 6.0 mg/ml, 12.0 mg/ml, 24.0 mg/fl), were shown to have significantly lower OD readings (p<0.05) when compared to yeast treated in lower concentrations of transferrin (1.5 mg/ml). The optical density (OD) of the ten strains of yeast were significantly lower (p<0.005) when treated with various concentrations of transferrin than with the saline control except at 72 hours post incubation. These results indicate that serum has inhibitory effects on Malassezia pachydermatis growth in vitro, and transfferin is one of the components that contribute towards this inhibitory role. The inhibitions are dose and time dependent.

Fermentation-Mediated Enhancement of Ginseng's Anti-Allergic Activity against IgE-Mediated Passive Cutaneous Anaphylaxis In Vivo and In Vitro

  • Hwang, Seon-Weon;Sun, Xiao;Han, Jun-Hyuk;Kim, Tae-Yeon;Koppula, Sushruta;Kang, Tae-Bong;Hwang, Jae-Kwan;Lee, Kwang-Ho
    • Journal of Microbiology and Biotechnology
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    • v.28 no.10
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    • pp.1626-1634
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    • 2018
  • Ginseng (the root of Panax ginseng Meyer) fermented by Lactobacillus plantarum has been found to attenuate allergic responses in in vitro and in vivo experimental models. Ginseng has been reported to also possess various biological functions including anti-inflammatory activity. The present study was aimed at comparing the anti-allergic effect of ginseng and fermented ginseng extracts on IgE-mediated passive cutaneous anaphylaxis in vitro in a murine cell line and in vivo in mice. Fermented ginseng extract (FPG) showed higher inhibitory effect against in vitro and in vivo allergic responses when compared with ginseng extract (PG). The secretion of ${\beta}$-hexosaminidase and interleukin (IL)-4 from the IgE-DNP-stimulated RBH-2H3 mast cells were significantly (p < 0.05) inhibited by FPG treatment, and this effect was concentration-dependent. Further, MKK4 activation and subsequent JNK phosphorylation were attenuated by FPG treatment. The inhibitory effect of FPG on the in vitro allergic response was verified in vivo against IgE-DNP-induced passive cutaneous anaphylaxis in a mouse model. These data indicated that the fermentation of ginseng with L. plantarum enhanced its anti-allergic effects both in vitro and in vivo. We predict that compositional changes in the ginsenosides caused by the fermentation may contribute to the change in the anti-allergic effects of ginseng. The results of our study highlight the potential of the use of FPG as a potential anti-allergic agent.

Effect of Zona Hardening on In Vitro Fertilization in Mouse Oocytes III. Analysis of In Vitro Fertilization and Zona Hardening in Oocytes Treated with Peroxidase Inhibitors and Tyrosine Analogue (생쥐난자에 있어서 투명대 경화현상이 체외수정에 미치는 영향 III. Peroxidase Inhibitors와 Tyrosine Analogue 처리된 난자의 투명대 경화 현상과 체외수정)

  • 이상진;정길생
    • Korean Journal of Animal Reproduction
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    • v.17 no.3
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    • pp.183-191
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    • 1993
  • These experiments were carried out to investigate whether the enzyme is involved in zona hardening during normal activatin of the oocytes by sperm, and demonstrate peroxidase activity during in vitro fertilization of oocytes treated with peroxidase inhibitors(250 $\mu$M phenylhydrazine, 28mM sodium sulfite, 350mM glycine ethyl ester and 50mM sodium azide) and tyrosine analogue(12.5mM tyramine). Also, zona soluble properties of the ovarian oocytes incubated for 0, 5, 10 and 15 hr in the presence of pheylhydrazine or tyramine were studied by using $\alpha$-chymotrypsin. The results obtained from these experiments were summarized as follows ; 1. The rates of fertilizatin in control oocytes and oocytes treated with phenylhydrazine or tyramine were 69.8%, 62.3% and 88.2%, respectively. However in vitro fertilization in oocytes treated with three different peroxidase inhibitors, sodium sulfite, glycine ethyl ester and sodium azide, were not induced. The oocytes treated with phenylhydrazine had no significant effect on in vitro fertilization rate as compared to control. However there was a significantly different in fertilization between tyramine treated group and control group(P<0.01). 2. The zona solubility(t50) of control and fertilized oocytes in culture treated with phenylhydrazine or tyramine were 30.7, 26.0 and 16.3 min., respectively. Phenylhydrazine treated group and tyramine treated group had effect on inhibition of zona hardening as compared to control group. These results suggest that ovoperoxidase is involved in zona hardening during normal activation of the oocytes by sperm. 3. t50 of control oocytes and ovarian oocytes treated with phenylhydrazine or tyramine for 5, 10 and 15 hr in vitro were 14.0, 26.2 and 32.0 min., 14.5, 26.9 and 30.2 min., and 14.0, 24.3 and 31.2 min., respectively. These results suggest that zona hardening in ovarian oocytes matured for various times in vitro cannot be inhibited by peroxidase inhibitors and tyrosine analogue, that the spontaneous zona hardening incultured ovarian oocytes is not caused by the secretory products of cortical granules released during the cortical reaction, ovoperoxidase.

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Inhibitory Effect of Methanol Extracts and Solvent Fractions from Doenjang on Mutagenicity Using in vitro SOS Chromotest and in vivo Drosophila Mutagenic System (된장 메탄올 추출물 및 분획물에 의한 in vitro SOS Chromotest 실험계와 in vivo 초파리 돌연변이 검출계에서의 항돌연변이 효과)

  • Lim, Sun-Young;Lee, Sook-Hee;Park, Keun-Young;Yun, Hee-Sun;Lee, Won-Ho
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.33 no.9
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    • pp.1432-1438
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    • 2004
  • This study investigated the inhibitory effect of methanol extracts and several solvent fractions from doen-jang on mutagenicity using in vitro SOS chromotest and in vivo Drosophila mutagenic system. In order to determine an antimutagenic effect of doenjang methanol extracts, other soybean fermented foods and original materials were compared. The treatment of doenjang methanol extracts (100 ${\mu}$/assay) to SOS chromotest system inhibited N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) induced mutagenicity by 87~97% and showed higher antimutagenic effect than other fermented foods. Among solvent fractions from doenjang methanol extracts, the ethylacetate and dichloromethane fractions showed the stronger antimutagenic effect (91% and 95%, respectively) in SOS chromotest. In Drosophila mutagenic system, the treatment of ethylacetate fraction (5%/bottle) significantly inhibited aflatoxin $B_1$ induced mutagenicity by 97%. These results demonstrated that doenjang had an inhibitory effect to mutagenic agents in both in vitro and in vivo mutagenic systems, suggesting that its antimutagenic effect may be due to active compounds in the ethylacetate fraction from doenjang methanol extracts.

In Vitro and In Vivo Physiological Characteristics of Dietary Fiber from By-product of Aloe vera Gel Processing (알로에 베라 유래 식이섬유의 In Vitro 및 In Vivo 생리기능 특성)

  • Baek, Jin-Hong;Cha, Tae-Yang;Heo, Jin-Chul;Lee, Sang-Han;Lee, Shin-Young
    • Food Engineering Progress
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    • v.14 no.2
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    • pp.173-182
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    • 2010
  • A fiber fraction (Aloe cellulose), the by-product obtained from Aloe vera gel processing was freeze dried and investigated for in vitro glucose/ bile acid retarding effects of powdered sample (100 mesh) comparing with commercial $\alpha$-cellulose as a reference sample. We also examined the effectiveness of physiological functionality such as the antiobesity and anti-constipation on Sprague-Dawley (SD) rat. The Aloe cellulose powders during in vitro dialysis experiment for 2 hours exhibited the glucose and bile acid retarding index of 20.32-35.2% and 53.13-28.30%, respectively. Especially, freeze dried aloe cellulose showed the 2.5 and 1.2-6 times higher effect on in vitro glucose and bile acid retardation than those of $\alpha$-cellulose. These relatively good retarding effects on glucose and bile acid diffusion suggest a potential of preventing from diabetes and arteriosclerosis of some extent. Also, the results from animal experiments on SD rats fed a high-fat diet for 4 weeks suggested that Aloe cellulose might be used as a novel dietary fiber showing an effective anti-obesity and anti-constipation effect.

Effect of Exocytosis Factor on Spontaneous Zona Pellucida Hardening during in Vitro Culture of the Mouse Oocytes (생쥐 난자 배양시 외분비 관련 요소들이 자발적 투명대 경화 현상에 미치는 영향)

  • Kang, Hye-Na;Bae, In-Ha;Kim, Hae-Kwon
    • Clinical and Experimental Reproductive Medicine
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    • v.21 no.1
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    • pp.111-119
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    • 1994
  • "Spontaneous" hardening of the zona pellucida of mouse oocytes during in vitro culture is most likely due to cortical granules exocytosis. Thus the purpose of the present study was to determine whether the exocytosis factor is involved in spontaneous zona pellucida hardening during in vitro culture of the mouse. The results obtained form these experiments were summarized as follows; 1. When a protein synthesis inhibitor(100${\mu}g$/ml puromycin) was added to the culture medium, it did not prevent spontaneous ZPH of mouse oocyte during in vitro culture. 2. Calmodulin antagonists (trifluoperazine and chlorpromazine) and calcium channel blocker (verapamil) had no inhibitory effect in spontaneous ZPH. 3. A microtubule assembly inhibitor, colcemid had some inhibitory effect on spontaneous ZPH. 4. Treatment with a microfillament formation blocker(cytochalasin-B) at 1${\mu}g$/ml concentration, resulted in the excellent inhibitory effect on spontaneous ZPH. However cytochalasin-B did not inhibit ethanol-induced ZPH.

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