Category: Vitro Fertilization

The viability of in vitro-derived embryos obtained from oocytes matured in the presence of FSP was evaluated by transferring 2- to 4-cell-stage embryos to recipient gilts. Of 9 recipients, 5 became pregnant and gave birth to 18 live piglets. Although 40 embryos were transferred to the oviduct of each recipient, the litter size ranged from 1 to 6 piglets, thus indicating low embryo survival. The exact rea-son(s) for the low embryo survival is not known, but a lack of proper interaction between embryos and the recipient cannot be ruled out. Mattioli et al. , Yoshida et al. , and Funahashi et al. also have reported the birth of piglets after transfer of IVM-IVF-derived embryos to recipient gilts. Those studies used freshly ejaculated boar semen for IVF. In the present study we used frozen-thawed ejaculated semen for IVF, and to our knowledge this is the first report to describe the successful establishment of pregnancies and birth of piglets using frozen-thawed semen for IVF. Although piglets were obtained after embryo transfer, the efficacy of IVM/IVF/IVP and/or embryo transfer techniques needs to be improved further in order to increase the average litter size if in vitro techniques are to be used as a profitable device for commercial applications. birth control pills

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DISCUSSION(4)

In the present study, the exact mechanism by which FSP influenced a higher GSH concentration in IVM pig oocytes is not known. It is possible that FSP may have prevented the oxidation of available cysteine in the maturation medium, a precursor of GSH, so that oocytes may effectively utilize cysteine to synthesize GSH. It has been shown that, outside the cell, the cysteine present in culture medium is unstable and readily oxidized to cystine within 1 h . Consequently, GSH synthesis may be impaired under such culture conditions. Only a small amount of cystine is taken up by denuded pig oocytes, which results in a low intracellular GSH concentration . We suggest that the presence of FSP during IVM may lower the oxygen tension, as reported for oviduct cells in culture , thus preventing the oxidation of cysteine to cystine. If this hypothesis is true, oocytes cocultured with FSP will have a higher amount of cysteine in the culture medium than oocytes cultured in medium without FSP. ventolin inhalers

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Improved development of 6- to 8-cell bovine embryos to the blastocyst stage is associated with increased intracellular GSH . Furthermore, a high intracellular concentration of GSH also appears to be essential for overcoming the in vitro developmental arrest in mice and rats . In the present study, intracellular GSH was significantly higher in pig oocytes cocultured with FSP during IVM and could be responsible for the improved blastocyst development. Although the relationship between higher intracellular GSH concentration and increased embryo development to the blastocyst stage is not clear, it is possible that embryos derived from fertilized oocytes with a low GSH content may be subjected to oxidative damage resulting in poor embryonic development. buy levaquin online

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DISCUSSION(2)

The embryo culture medium used in the present study was NCSU 23 medium supplemented with 0.4% BSA. Recent experiments with NCSU 23 medium provide evidence that in vitro development of pig embryos, derived from IVM oocytes fertilized in vivo or in vitro, can develop to morula and blastocyst stages . This medium could be used to predict the ability of embryos to develop prior to embryo transfer . To date there are only a few reports describing in vitro development of IVM-IVF pig embryos. Except in the two studies reported from our laboratory , in vitro development to the blastocyst stage (< 12%) has generally been extremely low . In the present study we were successful in obtaining a much higher (36%) blastocyst rate when IVM was carried out in the presence of FSP. Buy Asthma Inhalers Online

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The change of medium after 24 h is similar to our IVM conditions except for the differences in the type of maturation medium and supplements. According to Mattioli et al. , the process by which follicular somatic cell secretions influence complete oocyte maturation is due to the ability to maintain a functional intercellular coupling between cumulus cells and the oocyte. However, on the basis of their study, Liu et al. suggested that the full maturation of porcine oocytes in vitro is related to an intricate combination of actions of factors secreted by follicle cells and gonadotropic hormones. cialis professional

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RESULTS(2)

Effect of Coculture with FSP on Intracellular GSH Concentration

A total of six samples (20-30 oocytes per sample) from three replicates were assayed. As shown in Figure 4, the intracellular concentration of GSH in oocytes matured in the presence of FSP (8.7 ±1.6 pmol/oocyte) was significantly (p < 0.01) higher than in oocytes matured in the absence of FSP (6.4 ±1.8 pmol/oocyte).

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Embryo Transfer

Prepubertal gilts aged 165-180 days were used as recipients. These animals received a single i.m. injection of 1000 IU eCG followed by 500 IU hCG 84-96 h later. Embryos (2 to 4 cell, n = 40) derived 48 h after IVF of oocytes matured in the presence of FSP were surgically transferred into one oviduct each of 9 recipients 72 h after the hCG injection. Pregnancy diagnosis was carried out by an ultrasound scanner (Aloka 500V; Corometrics Medical Systems, Inc., Wallingford, CT) 30-35 days after the hCG injection, and pregnant animals were allowed to farrow. buy ampicillin

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