• Title, Summary, Keyword: In Vivo Development

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Embryo Gender Ratio and Developmental Potential after Biopsy of In Vivo and In Vitro Produced Hanwoo Embryos

  • Cho, Sang-Rae;Choe, Chang-Young;Son, Jun-Kyu;Cho, In-Cheol;Yoo, Jae-Gyu;Kim, Hyung-Jong;Ko, Yeong-Gyu;Kim, Nam-Young;Han, Sang-Hyun;Park, Yong-Sang;Ko, Moon-Suck
    • Reproductive and Developmental Biology
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    • v.36 no.4
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    • pp.269-273
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    • 2012
  • The present study was to assess the in vitro viability and sexing rate of bovine embryos. Blastocysts were harvested on day 7~9 day after insemination(in vitro and in vivo), and the sex of the embryos was examined using the LAMP method. Embryo cell biopsy was carried out in a $80{\mu}l$ drop $Ca^{2+}$, $Mg^{2+}$ free D-PBS and, biopsied embryos viability were evaluated after more 12 h culture in IVMD culture medium. The formation of recovered embryo to expanded and hatching stages had ensued in higher of sexed embryo in vivo than in vitro (100% vs. 89%, p<0.05), and in vitro, the rates of degeneration after sexing were significantly (p<0.05) higher in vitro than in vivo(11% vs. 0.0%). The rates of the predicted sex were female 61% vs. 56%, and male 39% vs. 44% in vivo and in vitro, respectively. The rates of survival following different biopsy methods were seen between punching and bisection method in vivo and in vitro (100% vs. 89% and 100% vs, 78% respectively). Biopsy method by punching was significantly (p<0.05) higher than bisection between produced embryos in vivo and in vitro. The present data indicate that with microblade after punching for embryo sexing results in high incidence of survivability on development after embryo biopsy. It is also suggested that LAMP-based embryo sexing suitable for field applications.

Development of Early Embryos inIn Vivo Superovulated Rabbits (과배란 처리된 체내 초기배 발생에 관한 연구)

  • 조현조;이홍준;심금섭
    • Journal of Embryo Transfer
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    • v.9 no.2
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    • pp.167-172
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    • 1994
  • This experiment was arried out to investigate the development of ea4y rabbit embryos in vivo. Twenty-six New Zealand White does were superovulated by treatment with PMSG(Intervet Co; I. M single injection, 150. U./rabbit) followed 3 day later by simultaneous I.V injection of 100 I.U HCG (Intervet Co, )and natural service with fertile male. All of does was killed at the specific times (24, 27, 30, 36, 42, 50 and 93 h post-hCG) to find out the early embryonic development in vivo respectively. Embryos at the specified stages of development were obtained at the following times after injection of hCG; one-ceH at 24 h, two-cell at 24~27h, four-cell at 27~36 h, morulae at 50 h and early blasto-cyst at 93 h and expanded or hatching blastocyst at 144 h. Number of embryos recovered per rabbit superovulated was 26.1 and average of recovery rate was 83.7%. The results suggest that superovulation was efficient for the increase of embryo number in rabbits, and as shown in results, asynchronous cleavage was prevalent among the recovered embryos.

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Adaptive Transition of Aquaporin 5 Expression and Localization during Preimplantation Embryo Development by In Vitro Culture

  • Park, Jae-Won;Shin, Yun Kyung;Choen, Yong-Pil
    • Development and Reproduction
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    • v.18 no.3
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    • pp.153-160
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    • 2014
  • Adaptive development of early stage embryo is well established and recently it is explored that the mammalian embryos also have adaptive ability to the stressful environment. However, the mechanisms are largely unknown. In this study, to evaluate the possible role of aquaporin in early embryo developmental adaptation, the expression of aquaporin (AQP) 5 gene which is detected during early development were examined by the environmental condition. To compare expression patterns between in vivo and in vitro, we conducted quantitative RT-PCR and analyzed localization of the AQP5 by whole mount immunofluorescence. At in vivo condition, Aqp5 expressed in oocyte and in all the stages of preimplantation embryo. It showed peak at 2-cell stage and decreased continuously until morula stage. At in vitro condition, Aqp5 expression pattern was similar with in vivo embryos. It expressed both at embryonic genome activation phase and second mid-preimplantation gene activation phase, but the fold changes were modified between in vivo embryos and in vitro embryos. During in vivo development, AQP5 was mainly localized in apical membrane of blastomeres of 4-cell and 8-cell stage embryos, and then it was localized in cytoplasm. However, the main localization area of AQP5 was dramatically shifted after 8-cell stage from cytoplasm to nucleus by in vitro development. Those results explore the modification of Aqp5 expression levels and location of its final products by in vitro culture. It suggests that expression of Aqp5 and the roles of AQP5 in homeostasis can be modulated by in vitro culture, and that early stage embryos can develop successfully by themselves adapting to their condition through modulation of the specific gene expression and localization.

Biosynthesized Platinum Nanoparticles Inhibit the Proliferation of Human Lung-Cancer Cells in vitro and Delay the Growth of a Human Lung-Tumor Xenograft in vivo -In vitro and in vivo Anticancer Activity of bio-Pt NPs-

  • Bendale, Yogesh;Bendale, Vineeta;Natu, Rammesh;Paul, Saili
    • Journal of Pharmacopuncture
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    • v.19 no.2
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    • pp.114-121
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    • 2016
  • Objectives: Lung cancer remains a deadly disease with unsatisfactory overall survival. Cisplatin, a standard platinum (Pt)-based chemotherapeutic agent, has the potential to inhibit the growth of lung cancer. Its use, however, is occasionally limited by severe organ toxicity. However, until now, no systematic study has been conducted to verify its efficacy with proper experimental support in vivo. Therefore, we examined whether biosynthesized Pt nanoparticles (NPs) inhibited human lung cancer in vitro and in vivo to validate their use in alternative and complementary medicine. Methods: We evaluated the in vitro and the in vivo anticancer efficiencies of biosynthesized Pt NPs in a subcutaneous xenograft model with A549 cells. Severe combined immune deficient mice (SCID) were divided into four groups: group 1 being the vehicle control group and groups 2, 3 and 4 being the experimental groups. Once the tumor volume had reached $70-75mm^3$, the progression profile of the tumor growth kinetics and the body weights of the mice were measured every week for 6 weeks after oral administration of Pt NPs. Doses of Pt NPs of 500, 1,000 and 2,000 mg/kg of body weight were administered to the experimental groups and a dose of honey was administered to the vehicle control group. The efficacy was quantified by using the delay in tumor growth following the administration of Pt NPs of A549 human-lung-cancer xenografts growing in SCID mice. Results: The in vitro cytotoxicity evaluation indicated that Pt NPs, in a dose-dependent manner, inhibited the growth of A549 cells, and the in vivo evaluation showed that Pt NPs at the mid and high doses effectively inhibited and delayed the growth of lung cancer in SCID mice. Conclusion: These findings confirm the antitumor properties of biosynthesized Pt NPs and suggest that they may be a cost-effective alternative for the treatment of patients with lung cancer.

ULTRASTRUCTURAL COMPARISON OF BOVINE BLASTOCYSTS DEVELOPED IN VIVO AND IN VITRO

  • Ohboshi, S.;Nakamichi, R.;Hanada, K.;Zhao, J.;Hattori, M.;Fujihara, N.;Umetsu, R.
    • Asian-Australasian Journal of Animal Sciences
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    • v.8 no.6
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    • pp.599-605
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    • 1995
  • The ultrastructures of in vitro-derived bovine blastocysts have been compared with those of blastocysts obtained from a superovulated cow. In vivo blastocysts obtained from the uterus showed well-differentiated features, while in vitro-derived embryos, which were developed from in vitro fertilized ovum, showed insufficient cellular organizations. In vitro-derived embryos contained many undefined cellular organizations in the perivitelline spaces compared with in vivo-derived blastocysts. Other features of in vivo and in vitro blastocysts were characterized by differential development of microvilli projection into blastocoele from the surface of the trophoblast cells. The conceivable reason for the difference between in vivo and in vitro developments of bovine embryos is that it is likely that in vitro culture system adopted in the present experiment may not be sufficient for better embryonic development.

Results of Embryo Transfer with Hanwoo Embryos Produced In-Vivo or In-Vitro to Holstein Cows as Recipients (체내 또는 체외에서 생산된 한우 수정란을 젖소 수란우에 이식한 결과)

  • Kim, Yong-Jun;Park, Hoon;Lee, Hae-Lee;Shin, Dong-Su;Jo, Sung-Woo;Kim, Yong-Su;Kim, Sue-Hee
    • Journal of Embryo Transfer
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    • v.23 no.3
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    • pp.167-175
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    • 2008
  • This study was performed to investigate the result that in-vivo or in-vitro embryos of Hanwoo cows were transferred to Holstein cows. Seventeen Hanwoo cows were used as donors for production of in-vivo embryos and fresh hanwoo in-vivo embryos were transferred to 1,150 Holsteins. And 2 embryos were transferred to 188 Holstein recipients to produce twin calves. Diagnosis on pregnancy was performed by rectal palpation at $60\sim90$ days after transfer. The pregnancy rate of Holstein recipients was 55.8% after transfer with Hanwoo in-vivo embryos and 38.2% after transfer with Hanwoo in-vitro embryos. The delivery rate of pregnant Holstein recipients was 88.4% after transfer with Hanwoo in-vivo embryos and 75.6% after transfer with Hanwoo in-vitro embryos. The rate of delivery of Holstein recipients transferred with two Hanwoo embryos was 36.2% and the rate of twin production was 25.9%. The rate of twin production by embryo transfer with in-vivo embryos was 30.4%, whereas the fate with in-vitro embryos was 15.6%. The pregnancy rate according to the grade of corpus luteum of Holstein recipients transferred with Hanwoo in-vitro embryos was 41.5 and 36.0% for A and B grade, respectively. The pregnancy rate according to the transfer in site in the uterine lumen of recipients was 40.9 and 32.7% for anterior and middle site, respectively. The pregnancy rate according to day of embryo transfer after estrus of recipients was 45.5, 38.8 and 39.7% for day 6, day 7 and day 8, respectively. There was difference of pregnancy rate according embryo transfer technician ($30.5\sim45.8%$) individual dairy farm ($21.1\sim51.0%$). These results are supposed to indicate that the rate of pregnancy after transfer with Hanwoo embryos to Holstein recipients was similar to that within the same breed, and consequently that this method would be beneficial to enhance the productivity in Hanwoo reproduction.

Effects of Iron, Chelators and Nitrate Concentration on in vivo Fluorescence and Nitrate Reductase of the Red Tide Organism Amphidinium carterae

  • Yang, Sung-Ryull;Song, Hwan-Seok;Pae, Se-Jin;Huh, Sung-Hoi
    • Journal of the korean society of oceanography
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    • v.34 no.1
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    • pp.49-57
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    • 1999
  • A red tide organism, Amphidinium carterae was incubated under different iron/chelator and nitrate concentrations to investigate the factors controlling the growth. The chelation capacity played a critical role in regulating the nitrate reductase (NR) activity and in vivo fluorescence of this organism. However, there was a significant difference between the NR activity and in vivo fluorescence in response to trace metals and chelator treatments. In vivo fluorescence was the highest in FeEDTA 10 ${\mu}$M treatments and the lowest in DTPA 10 ${\mu}$M treatments. This indicates that the availability of the trace metal is important in regulating the in vivo fluorescence of this photosynthetic microalgae In contrast, NR activity showed the highest values in trace metal enriched treatments, and trace metal + DTPA treatments showed fairly high NR activities. This suggests that DTPA treatment did not hinder the NR activity as much as it did in vivo fluorescence. In vivo fluorescence and NR activity increased with nitrate concentration of up to 50 ${\mu}$M and remained relatively constant or the rate of increase decreased above that concentration, indicating that initial nitrate concentration of higher than a certain level would not accelerate the growth of A. carterae. Further investigation is needed to elucidate the reason for the difference in timing sequence between the NR and in vivo fluorescence in response to different metal treatments and chelation capacity.

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Properties of Two Cellular Biomarker Parameters in the Blood of Farmed Pacific Oyster, Crassostrea gigas, Exposed to Polychlorinated Biphenyls

  • Choy Eun Jung;Jo Qtae;Do Jeong Wan;Kim Sang Soo;Jee Young-Ju;Min Kwang Sik
    • Fisheries and aquatic sciences
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    • v.6 no.2
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    • pp.74-80
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    • 2003
  • Two cellular biomarker parameters of the farmed Pacific oyster Crassostrea gigas were studied in vivo and in vitro after exposure to concentrations of polychlorinated biphenyls in terms of neural red uptake (NRU) and lysozyme activity. The oysters exposed in vivo to the xenobiotic concentrations, 0, 30, 90, and 180 ng/g for 14 days, enhanced hemocyte NRU with occasional significant differences (P<0.05), depending on the chemical concentration and duration. An adverse tendency was manifest in the lysozyme activities both in the hemocyte and serum of the oyster treated with the chemical in a same manner, rendering these two cellular parameters as biomarker candidates against the chemical. The oysters exposed in vitro to the chemical concentrations, 0, 1, 5, 10, 100, 1,000, and 10,000 ng/g for 24 hrs at $10^{\circ}C$ showed a similar tendancy as those exposed in vivo to the chemical. Unlike in vivo response, however, the in vitro NRU was first influenced by very low concentration of the chemical. In in vitro results, marked but not significant increase of hemocyte NRU was noticed at the chemical concentration of 5 ng/g, where the value was almost as high as those exposed to higher chemical concentrations, up to 10,000 ng/g. An unusual result was observed in the in vitro lysozyme activity of hemocyte in which significant decrease was first noticed at the chemical concentration of 100 ng/g.

Ex vivo Cytotoxicity of the Bacillus thuringiensis Cry4B δ-Endotoxin to Isolated Midguts of Aedes aegypti Larvae

  • Barusrux, Sahawat;Sramala, Issara;Katzenmeier, Gerd;Bunyaratvej, Ahnond;Panyim, Sakol;Angsuthanasombat, Chanan
    • BMB Reports
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    • v.36 no.3
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    • pp.294-298
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    • 2003
  • The pathological effect of the Bacillus thuringiensis Cry $\delta$-endotoxins on susceptible insect larvae had extensive damage on the midgut epithelial cells. In this study, an ex vivo assay was devised for assessing the insecticidal potency of the cloned Cry4B mosquito-larvicidal protein that is expressed in Escherichia coli. Determination of toxicity was carried out by using a cell viability assay on the midguts that were dissected from 5-day old Aedes aegypti mosquito larvae. After incubation with the toxin proteins, the number of viable epithelial cells was determined photometrically by monitoring the quantity of the bioreduced formazan product at 490 nm. The results showed that the 65-kDa trypsin-activated Cry4B toxin exhibited toxic potency ca. 3.5 times higher than the 130-kDa Cry4B protoxin. However, the trypsin-treated products of the non-bioactive Cry4B mutant (R158A) and the lepidopteran-specific Cry1Aa toxin displayed relatively no ex vivo activity on the mosquito-larval midguts. The ex vivo cytotoxicity studies presented here confirms data that was obtained in bioassays.

The Emergence of Behavioral Testing of Fishes to Measure Toxicological Effects

  • Brooks, Janie S.
    • Toxicological Research
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    • v.25 no.1
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    • pp.9-15
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    • 2009
  • Historically, research in toxicology has utilized non-human mammalian species, particularly rats and mice, to study in vivo the effects of toxic exposure on physiology and behavior. However, ethical considerations and the overwhelming increase in the number of chemicals to be screened has led to a shift away from in vivo work. The decline in in vivo experimentation has been accompanied by an increase in alternative methods for detecting and predicting detrimental effects: in vitro experimentation and in silico modeling. Yet, these new methodologies can not replace the need for in vivo work on animal physiology and behavior. The development of new, non-mammalian model systems shows great promise in restoring our ability to use behavioral endpoints in toxicological testing. Of these systems, the zebrafish, Danio rerio, is the model organism for which we are accumulating enough knowledge in vivo, in vitro, and in silico to enable us to develop a comprehensive, high-throughput toxicology screening system.