Effect of Replacement of Pyruvate/Lactate in Culture Medium with Glucose on Preimplantation Development of Porcine Embryos In Vitro

Karja, Ni Wayan Kurniani; Medvedev, Sergey; Ōnishi, Akira; Fuchimoto, Daiichiro; Iwamoto, Masao; Otoi, Takeshige; Nagai, Takashi

doi:10.1262/jrd.50.587

Cited by 13 publications

(23 citation statements)

References 22 publications

(39 reference statements)

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“…Abnormal cell division was also observed in porcine embryos produced by IVF at the first cleavage division (unpublished our data). The average number of cells per blastocyst found in the present study was similar to that previously reported by Kikuchi et al [8], Somfai et al [33] and Karja et al [34]. Blastocysts derived from 3-to 4-cell embryos and 5-to 8-cell embryos had significantly more cells than those from the other cell stage embryos.…”

Section: Discussionsupporting

confidence: 92%

“…In the present study, the overall blastocyst rate (23.1%) for all the inseminated oocytes was similar to the rate previously reported by Kikuchi et al [8] (25.3%); however, it was slightly lower than the rates reported by Somfai et al [33] (32%) and Karja et al [34] (31.3%). When the embryos were classified by their cleaved stages on Day 2, the blastocyst rate of the moderate stage embryos (3-to 4-cell and 5-to 8-cell embryos) was significantly higher than the rates of the delayed or slow (2-cell embryos) and fast stage (>8-cell embryos) embryos.…”

Section: Discussionsupporting

confidence: 90%

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The Blastocyst Production Rate and Incidence of Chromosomal Abnormalities by Developmental Stage in In Vitro Produced Porcine Embryos

Ullo

Yoshizawa

Komoriya

et al. 2008

Journal of Reproduction and Development

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Abstract. The present study was conducted to determine the relationship between embryonic development speed at different stages (the cleaved stage at 52 h and the blastocyst stage at 6 days post insemination) and incidences of chromosome abnormalities in in vitro produced porcine embryos. Porcine oocytes were collected from 3-6-mm ovarian follicles obtained at a slaughterhouse and matured in modified NCSU-37 medium for 44-46 h. Following in vitro fertilization with a final concentration of 1 × 10 5 sperm/ml for 3 h, all oocytes were cultured in vitro for 52 h. Day-2 (52 h after insemination) embryos were classified according to their cleaved stages into 2-cell, 3-to 4-cell, 5-to 8-cell, and >8-cell stages; these were cultured separately for additional 4 days (Day 6). The resultant Day-6 blastocysts were classified according to the morphological diameter into 3 grades: Grade A, expanded blastocysts; Grade B, expanding blastocysts; and Grade C, early blastocysts. They were then analyzed chromosomally. The 3-to 4-cell and 5-to 8-cell embryos had significantly high blastocyst development rates (46.1 and 36.9%, respectively), and these blastocysts contained significantly more cells (40.2 and 42.4 cells, respectively) than those derived from 2-cell embryos and >8-cell embryos (28.6 and 26.5 cells, respectively). The incidence of chromosomal abnormalities was significantly higher in the blastocysts derived from 2-cell and >8-cell stage embryos than in the blastocysts derived from the other stage embryos. Furthermore, the grade A blastocysts had the lowest incidence of chromosomal abnormalities (35.3%) and contained the most cells (48.7 cells). Porcine in vitro production (IVP) yielded a high blastocyst rate and an excellent embryo quality when 3-to 4-cell and 5-to 8-cell stage embryos were selected on Day 2 after insemination. The same criteria yielded a higher quality of expanded blastocysts based on the stage of embryo development and morphology. Key words: Blastocyst, Chromosomal abnormality, Embryo, In vitro production (IVP), Pig (J. Reprod. Dev. 54: [22][23][24][25][26][27][28][29] 2008) n mammals, the technology of in vitro production (IVP) of embryos, which involves in vitro maturation and fertilization of oocytes and in vitro culture of the resultant embryos, has become important for increasing the number of offspring from selected high-quality animals and for reducing the generation intervals for breed improvement in livestock [1,2]. Moreover, IVP technology has been used in the production of cloned and transgenic animals, which can be used in biomedical research [3][4][5]. In swine, although embryos have been successfully produced using the IVP system, the development rates to the blastocyst stage and to full term after embryo transfer are still low [6][7][8]. The viability of porcine embryos produced by IVP is low compared with that of in vivo-derived porcine embryos. For example, porcine embryos produced by IVP are of poor quality in terms of the number of cells compared with those produced in vivo [9].A...

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Section: Discussionsupporting

confidence: 92%

Section: Discussionsupporting

confidence: 90%

The Blastocyst Production Rate and Incidence of Chromosomal Abnormalities by Developmental Stage in In Vitro Produced Porcine Embryos

Ullo

Yoshizawa

Komoriya

et al. 2008

Journal of Reproduction and Development

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“…An alternative is to use embryos produced entirely in vitro. Obviously, a critical component in generating in vitro embryos is the culture medium in which they are grown.Several reports have investigated improving two commonly used porcine embryo culture media, North Carolina State University 23 and 37 (NCSU23 and NCSU37;[2]) [3][4][5]. As originally formulated, these media contain only glucose as the primary energy substrate.…”

mentioning

confidence: 99%

“…Karja et al [4] found the optimum time to change modified NCSU37 supplemented with pyruvate and lactate to modified NCSU37 supplemented with glucose was 58 h after the start of culture. However, the optimum time to change the energy substrates and add EAA has not yet been adequately investigated with porcine embryos in our modified NCSU23.…”

mentioning

confidence: 99%

Adding Essential Amino Acids at a Low Concentration Improves the Development of In Vitro Fertilized Porcine Embryos

Beebe

Vassiliev

McIlfatrick

et al. 2009

Journal of Reproduction and Development

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Abstract. The aims of this study were to investigate improvements to the pig preimplantation embryo culture system using in vitro produced embryos. For experiment 1, the optimum time to change the medium from NCSU23 containing 0.6 mM glucose, 0.2 mM pyruvate, 5.7 mM lactate and nonessential amino acids to NCSU23 containing 5.6 mM glucose and both essential and nonessential amino acids was examined. There were no statistically significant differences in blastocyst rates or cell number when the medium was changed at 48, 72 or 96 h, although there was a consistent trend for the 96 h treatment to produce fewer blastocysts with fewer cells. For experiment 2, the addition of essential amino acids at either a 1:50 or a 1:100 dilution of the purchased stock solution for day 1 to 6 or for days 3 to 6 only was investigated. Adding essential amino acids at a 1:50 dilution for day 3 to 6 significantly reduced the blastocyst rate and adding them at a 1:50 dilution from day 1 to 6 significantly reduced both the blastocyst rate and blastocyst cell number compared to when it was added at a 1:100 dilution. Embryos were produced by IVF, cultured for 6 days and good quality blastocysts were transferred into 6 synchronized pseudopregnant recipients (24 to 35 blastocysts per recipient) resulting in 4 pregnancies and 21 live birth piglets. These results show that adding essential amino acids at a 1:100 dilution provided the best culture conditions and the blastocysts produced were able to attain full term development after transfer. Key words: Blastocysts, Essential amino acids, In vitro culture, Piglets, Porcine (J. Reprod. Dev. 55: [373][374][375][376][377] 2009) he pig is becoming increasingly important as a biomedical research tool [1] and shows great promise as a model for embryonic stem cell technologies. The capacity to reliably generate embryonic stem cell lines is essential to using the pig as such a model. While in vivo produced embryos are ideal for producing cell lines, collecting them is expensive and time consuming. An alternative is to use embryos produced entirely in vitro. Obviously, a critical component in generating in vitro embryos is the culture medium in which they are grown.Several reports have investigated improving two commonly used porcine embryo culture media, North Carolina State University 23 and 37 (NCSU23 and NCSU37;[2]) [3][4][5]. As originally formulated, these media contain only glucose as the primary energy substrate. These reports found that providing pyruvate and lactate for the early preimplantation period (approximately 48 to 73 h post insemination) and only glucose for the remainder results in improved development and embryonic quality.Amino acids have been grouped by Eagle [6] into essential (EAA) and non-essential (NEAA), and are normally added to culture media as prepared concentrated stock solutions at a dilution of 1:50 for EAA and 1:100 for NEAA. Early cleavage development was found to be stimulated by the addition of NEAA to the culture medium but inhibited by EAA in both the mouse [...

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“…Several reports have investigated improving two commonly used porcine embryo culture media, NCSU-23 and 37 (Raychoudhury and Suárez, 1991;Kikuchi et al, 2002;Karja et al, 2004;Beebe et al, 2009). Very few experiments have studied the use of hormones in the maturation of porcine oocytes (Eroglu, 1993;Bing et al, 2001;Dode and Graves, 2002).…”

Section: Discussionmentioning

confidence: 99%

Progesterone improves porcine in vitro fertilisation system

Malo

Gil

Cano

et al. 2014

Acta Veterinaria Hungarica

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In an effort to improve the quality of in vitro produced porcine embryos, the effect of progestagens -progesterone analogues -on the in vitro developmental competence of porcine oocytes was studied. A total of 1421 in vitro matured oocytes, from 4 replicates, were inseminated with frozen-thawed spermatozoa. Progestagens were added to late maturation and embryo cultures (10 IU/ml). Fertilisation success (pre-maturation, penetration, monospermy and efficiency) and nuclear maturation were evaluated. There were no differences among prematuration rates between groups (P = 0.221). Penetration rates were higher (P < 0.001) in the presence of progestagens (75.0%) as compared to the control (51.7%). However, no differences were observed in monospermy percentages (P = 0.246). The results indicated that supplementation with progestagens increased the efficiency of the in vitro fertilisation system (P < 0.001). An additional beneficial effect was observed in nuclear maturation with progestagens (P = 0.035). In summary, progestagen supplementation is an important factor to improve the in vitro fertilisation procedure. Key words: In vitro fertilisation, oocyte maturation, progesterone, porcineThe overall efficiency of in vitro porcine embryo production and the quality of embryos are still low when compared with in vivo results (Funahashi, 2003). One of the major problems includes improper in vitro maturation (IVM) of oocytes, in both the nuclear and cytoplasmic compartments (Niwa, 1994;Marchal et al., 2001). While a large percentage of oocytes reached metaphase II after maturation, inadequate cytoplasmic and molecular maturation occurred (Sirard et al., 2006), leading to a lack of subsequent embryo development success. There is a lack of information on ovarian factors that may be important for the maturation and subsequent fertilisation of oocytes.The production of sex steroids by follicular cells is proposed to be influenced by the maturity of oocytes. Oestradiol, progesterone and testosterone are the main steroid hormones that play an essential role during the follicular and

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Effect of Replacement of Pyruvate/Lactate in Culture Medium with Glucose on Preimplantation Development of Porcine Embryos In Vitro

Cited by 13 publications

References 22 publications

The Blastocyst Production Rate and Incidence of Chromosomal Abnormalities by Developmental Stage in In Vitro Produced Porcine Embryos

The Blastocyst Production Rate and Incidence of Chromosomal Abnormalities by Developmental Stage in In Vitro Produced Porcine Embryos

Adding Essential Amino Acids at a Low Concentration Improves the Development of In Vitro Fertilized Porcine Embryos

Progesterone improves porcine in vitro fertilisation system

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