A single medium supports development of bovine embryos throughout maturation, fertilization and culture

Gandhi, Agharnan; Lane, Michelle; Gardner, David K.; Krisher, Rebecca L.

doi:10.1093/humrep/15.2.395

Cited by 75 publications

(62 citation statements)

References 48 publications

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“…*Calculated as a percentage of activated oocytes. medium can be compared simultaneously with minimal variation [14].…”

Section: Discussionmentioning

confidence: 99%

“…The IVP system using a single medium throughout IVM, IVF and IVC may be easier to use and maintain compared with media employed in standard IVP systems. In cattle, oocytes can be matured, fertilized and cultured in a single basic medium; namely, modified synthetic oviductal fluid (mSOF) appropriately supplemented for specific stages of development [14].The objective of the present study was to establish a defined system for IVP of porcine blastocysts. Moreover, we attempted to determine whether media based on the composition of porcine oviductal fluid could support IVP of porcine blastocysts through IVM, IVF and IVC.…”

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confidence: 99%

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confidence: 99%

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Defined System for In Vitro Production of Porcine Embryos Using a Single Basic Medium

2008

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Abstract. We have previously indicated that porcine blastocysts can be produced by in vitro fertilization (IVF) and culture (IVC) in chemically defined porcine gamete medium (PGM) and porcine zygote medium (PZM)-5, respectively, In the present study, the effects of basic media and macromolecular components on in vitro maturation (IVM) were investigated to develop a defined system for in vitro embryo production using a single basic medium through IVM, IVF and IVC. Porcine immature oocytes were matured in porcine oocyte medium (POM) or modified North Carolina State University (mNCSU) 37, which were supplemented with either 10% (v/v) porcine follicular fluid (pFF) or 3 mg/ml polyvinyl alcohol (PVA) as a macromolecular component (designated POM+pFF, POM+PVA, mNCSU37+pFF and mNCSU37+PVA). In the maturation with mNCSU37+PVA, the percentages of oocytes that reached the metaphase II stages were significantly lower than those in the other treatments. Following IVM with the above media, oocytes were treated with an electrical stimulus and cycloheximide for parthenogenetic activation and were cultured in PZM-5 for 5 days. The rates of cleavage and blastocyst formation of parthenogenetic oocytes were significantly lowered for maturation with mNCSU37+PVA compared with the other treatments, while there were no significant differences in the total numbers of cells in blastocysts among the treatments. Following IVF and IVC, the rates of penetration, male pronucleus formation, cleavage and blastocyst formation were significantly lower when oocytes were matured in mNCSU37+PVA than in other maturation media. The normal fertilization rate was significantly higher in POM+PVA compared with the other treatments, although the total number of cells in blastocysts was reduced with the addition of PVA to both POM and mNCSU37 compared with pFF supplementation. These results demonstrate that porcine blastocysts can be produced by the defined system using a single basic medium. Key words: Defined system, In vitro production (IVP), Maturation, Porcine embryo, Porcine zygote medium (PZM) (J. Reprod. Dev. 54: [208][209][210][211][212][213] 2008) he establishment of an in vitro production (IVP) system for preimplantation embryos that can develop to full term after transfer will contribute to a better understanding of the physiology of embryonic development in early pregnancy and the control of animal reproduction, including embryo transfer, transgenesis and cloning [1]. Numerous studies have investigated the ability of porcine oocytes and embryos to develop in vitro using a wide variety of culture media [2][3][4][5][6][7]. Recently, we developed a chemically defined medium (porcine zygote medium: PZM) for in vitro culture (IVC) of porcine zygotes based on the composition of pig oviduct fluid [8]. Moreover, we established a chemically defined system for in vitro fertilization (IVF) of porcine in vitro-matured oocytes using porcine gamete medium (PGM), modified from PZM, as a basic medium [9], and the in vivo viability of blastocysts produced i...

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“…*Calculated as a percentage of activated oocytes. medium can be compared simultaneously with minimal variation [14].…”

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

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Defined System for In Vitro Production of Porcine Embryos Using a Single Basic Medium

2008

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“…Addition of FBS or BSA has been extensively studied in bovine IVF during maturation or embryo culture (Fukui and Ono, 1989;Gandhi et al, 2000), but little is known regarding how protein type may affect bovine oocytes during fertilization.Thus, in the present study, we wanted to elucidate if prolonged incubation of IVM bovine oocytes in TCM supplemented with 10% FBS or 7 mg/ml of BSA differently affected the oocyte´s chromatin in the bovine species.…”

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confidence: 99%

Fetal bovine serum is associated with polar body degeneration after in vitro maturation of bovine oocytes

Macías‐García

Macedo

Rocha

et al. 2018

J Hellenic Vet Med Soc

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In vitro fertilization (IVF) in cattle is commonly used worldwide. Although extensive research has been conducted using different additives in the different IVF steps, little is known regarding how protein type may affect bovine oocytes during the fertilization period. In addition, unlike Tissue Culture Medium 199 (TCM), fertilization medium may induce oocytes’ chromatin degeneration during prolonged incubation in the horse (Modified Whitten’s medium). Thus, in the present work TCM-199 supplemented with either 7 mg/ml of Bovine Serum Albumin (TCM+BSA) or 10% Fetal Bovine Serum (v/v; TCM+FBS) was used. Bovine oocytes were matured in vitro and placed in the previously mentioned media for further 18 hours, in the absence of added sperm (sham fertilization) and their chromatin conformation was evaluated. After IVM, 78.9% of the initial oocytes had reached the MII stage. After sham fertilization, 58.6% of the oocytes in TCM+BSA while just 28.3% in TCM+FBS maintained the MII chromatin conformation (p < 0.05). Subsequent experiments run using PB extruded oocytes and incubated in TCM+BSA and TCM+FBS during sham fertilization, demonstrated that FBS was consistently associated with polar body dissolution or degeneration.

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“…An optimized IVP system using a single basic medium throughout IVM, IVF and IVC might be easier to use and maintain compared with media employed in standard IVP systems. In cattle, oocytes can be matured, fertilized and cultured in mSOF appropriately supplemented for specific stages of development [53]. Porcine oocyte medium (POM) is a modified PZM [54].…”

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confidence: 99%

Development and Application of a Chemically Defined Medium for the In Vitro Production of Porcine Embryos

Yoshioka

2011

J. Reprod. Dev.

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Abstract. Recent advances in systems for in vitro production (IVP) of porcine embryos, including in vitro oocyte maturation, fertilization and embryo culture, have enabled us to generate viable embryos that can develop to full term after transfer into recipients. This technology is being applied now to developments in gamete/embryo biology and agriculture, as well as in producing cloned and genetically modified pigs. Chemically defined media for IVP of embryos are useful for a precise analysis of the physical action of substances on gametogenesis and early embryogenesis, because they eliminate undefined factors present in biological materials, such as serum or serum albumin. Use of a chemically defined medium also improves the reliability of media formulations, yields a higher reproducibility of results and ensures biosafety of culture media by eliminating protein preparations, which may be contaminated with pathogens. Therefore, it has certain advantages for research and for commercial purposes. We have recently developed a defined IVP system for porcine embryos using a single basic medium based on the composition of porcine oviductal fluid. This paper discusses the developmental ability and normality of porcine IVP embryos, and limitations and advancements in this system. Key words: Chemically defined medium, Developmental competence, In vitro production, Piglet (J. Reprod. Dev. 57: [9][10][11][12][13][14][15][16] 2011) standard in vitro production (IVP) system for preimplantation embryos includes three technological steps: the in vitro maturation (IVM) of immature oocytes, in vitro fertilization (IVF) and in vitro culture (IVC) of zygotes. Successful large-scale IVP of porcine embryos will enable us to reduce the cost and time required and will be valuable for research in reproductive physiology, agriculture and biotechnology. The first successful production of piglets from in vitro-matured and in vitro-fertilized oocytes was reported by Mattioli et al. in 1989 [1], in which 2-to 4-cell embryos at 44 h after IVF were transferred into recipients. Since then, some laboratories have succeeded in producing piglets from cleaved embryos at the 2-to 4-cell stage cultured for 24-36 h [2][3][4] and from 8-cell to morula-stage embryos cultured for 96 h [5] following IVM and IVF. Moreover, in the present decade it has been demonstrated that porcine in vitro-produced blastocysts cultured for 5 or 6 days after IVF could develop to full term [6,7]. However, despite recent improvements in porcine IVP techniques, the developmental rate of embryos matured and fertilized in vitro to the blastocyst stage and their quality are still low compared with in vivo-derived embryos [8]. The low developmental competence of porcine in vitro-produced embryos might be caused by several factors, including a reduced incidence of male pronuclear formation, a high incidence of polyspermy and suboptimal conditions for embryo culture [9,10].Numerous media, such as modified Whitten's medium [11], North Carolina State University (NCSU) 23 medium...

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A single medium supports development of bovine embryos throughout maturation, fertilization and culture

Cited by 75 publications

References 48 publications

Defined System for In Vitro Production of Porcine Embryos Using a Single Basic Medium

Defined System for In Vitro Production of Porcine Embryos Using a Single Basic Medium

Fetal bovine serum is associated with polar body degeneration after in vitro maturation of bovine oocytes

Development and Application of a Chemically Defined Medium for the In Vitro Production of Porcine Embryos

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