227 resultados para In vitro production


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Pós-graduação em Biopatologia Bucal - ICT

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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

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This study was carried out to assess the influence of bovine embryo culture medium Beltsville Agriculture Research Center (BARC), supplemented with FCS, BSA or PVA, on the in vitro oocyte maturation, evidenced by cleavage rate and blastocysts production at different developmental stages. Three experiments were performed, as follows: exp.1: addition of FCS to BARC medium at concentrations of 0, 5 and 10%; exp. 2: addition of BSA to BARC medium at concentrations of 0, 4 and 8 mg/ml; exp. 3: addition of PVA to BARC medium at concentrations of 0, 0.5 and 1.0 mg/ml. TCM 199 supplemented with bicarbonate, pyruvate, gentamicin sulfate, FSH, LH and FCS was used as control group. Oocytes obtained from cow ovaries at slaughterhouse were selected in PBS, and then matured in BARC medium supplemented with FSH, LH and gentamicin sulfate, according to the experimental design. Percoll gradient was used for sperm selection and TALP medium for IVF. In vitro embryo culture was in SOF-m medium; a humidified atmosphere with 5% CO2, in air, at 38.7oC was used for all steps. The number of oocytes reaching blastocyst, expanded blastocyst, and hatched blastocyt stages was recorded, respectively at 72 and 168 h post-insemination. ANOVA and Bonferroni t test were used to determine differences among groups. Differences of P<0.05 were taken as significant. Higher percentage (P<0.05) of cleaved oocytes was observed in group TCM + FCS than for the other groups matured in BARC supplemented with FCS or BSA, regardless the concentration used. However, the cleavage rate was similar between groups BARC plus PVA with 1 mg/ml (85.7%) and TCM + FCS (90.8%). Significant difference was found among groups for the production of blastocysts, with the control group yielding a higher number of blastocysts (results ranging from 47.4 to 51.4%, in comparison with groups using BARC + FCS (4.1 to 19.7%), BSA (1.4 to 5.6%) and PVA (5.7 to 10.6%). In conclusion, BARC medium supplemented with different macromolecules did not promote a beneficial effect on in vitro oocyte maturation, resulting in lower rate of cleavage and blastocyst production when compared with TCM + FCS medium.

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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

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Pós-graduação em Agronomia (Produção Vegetal) - FCAV

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Despite the efforts made to improve the production of bovine embryos in vitro, their efficiency is still low, since only 30-40% of developed blastocysts are obtained from oocytes after in vitro maturation (IVM), fertilization and cultured embryos. Assisted reproductive technologies have a limiting impact due a lack of oocytes capable to fertilization.The comprehension of mechanism involved in oocyte maturation are crucial to establish a culture system that allows a larger number production of good quality embryos. The study of the early stages of oocyte and follicle development in vivo is important for a better understanding of the molecular pathways that regulate oogenesis, folliculogenesis and oocyte maturation. Thus the physiological biochemical and molecular mechanisms involved in maturation may contribute to the increased efficiency of in vitro embryo production. Therefore, the aim of this literature review is to understand the basic mechanisms that underlie oocyte maturation in cattle, since oocyte and follicle cells in vivo formation to its use in the in vitro environment.

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Lentinus strigosus (Schwein.) Fr. is an exploitable edible mushroom occurring in the Brazilian Amazon, being part of a huge diversity of edible mushrooms which are little grown. The use of regional waste is recommended to reduce production costs of any kind of edible mushroom. Thus, the mycelial growth of L. strigosus in culture media based on regional wood waste extract by using substrates based on Protium puncticulatum, Cariniana micrantha and Caryocar glabum sawdust, supplemented with 20% of wheat bran (Triticum aestivum), corn bran (Zea sp.) or rice bran (Oryza sp.) was observed. Eucalyptus (Eucaliptus sp.) sawdust was used for comparison with the other wood wastes because it is commonly used in the cultivation of edible fungi. The experimental design employed was totally randomized, in 4 x 3 factorial scheme (sawdust x bran), adding up 12 treatments with 5 repetitions, being that each repetition corresponded to a Petri dish, totalizing 60 dishes, incubated at 35 ºC. The diameter of the colony was daily evaluated until the fungus reached the borders of the Petri dish in one of the treatments. After that period, the media based on P. puncticulatum sawdust obtained thebest results of mycelial growth, showing potential to be used as an alternative residuein a future production of L. strigosus in the state of Amazonas.

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Aims: This work aimed to assess how seed size, vials, vial sealing (in vitro), and substrate (in vivo) affect C. regium germination and emergence. This study shall contribute to the viable production of C. regium seedlings. Study Design: The experimental design used in these experiments was randomized. Place and Duration of Study: Department of Plant Biotechnology, Universidade de RibeirãoPreto, between March 2010 and December 2010. Methodology: This work has evaluated how seed size, vials, vial sealing (in vitro), and substrate (in vivo) influence the germination and emergence of C. regium. Results: The results showed that cultivation of C. regium seedlings from seeds is viable, irrespective of seed size. Vial oxygenation is an important parameter to consider in vitro, to obtain a larger number of normal seedlings. As for in vivo conditions, germination should be conducted in sand, to ensure a greater amount of young seedlings. Conclusion: The results presented here attested that it is possible to produce C. regium seedlings from seeds of any size both in vivo and in vitro conditions. In vitro, it is important to consider vial oxygenation, in order to obtain a greater amount of normal seedlings. In vivo, germination should be conducted in sand, to ensure production of a large quantity of seedlings.