Effect of glucose levels during the in vitro culture in synthetic oviduct fluid medium on in vitro development of bovine oocytes matured and fertilized in vitro
“…Among other components investigated during in vitro maturation, fertilization, and culture (IVMFC) are sodium bicarbonate (NaHCO 3 ) [32,33]; insulin, transferrin, and selenium [34]; growth factors [35,36]; gas atmosphere [37]; glucose [38]; penicillamine (P) [39]; antioxidants [40]; and water quality [41]. However, completely defined conditions have been unavailable for definitive testing; either serum or BSA has been included for one or more of the steps composing the IVMFC procedure.…”
The objective was to establish an in vitro system in which bovine oocytes can be matured, fertilized, and cultured up to the blastocyst stage without support of serum, BSA, or somatic cells. Media consisted of modified tissue culture medium 199 (mTCM 199) with ovine LH (oLH) for maturation (IVM), experimental alterations of modified defined medium (mDM) for sperm selection and insemination (IVF), and citrate+synthetic oviductal fluid+nonessential amino acids (c-SOF+NEA) for zygote/embryo culture (IVC). Effects of heparin, BSA, polyvinyl alcohol (PVA), penicillamine (P), Hepes, and sodium bicarbonate (NaHCO3) were studied. Results included proportions of oocytes that cleaved by 48 h and that reached morulae by 120 h, blastocysts by 168 h, and expanded blastocysts by 216 h postinsemination (pi). Best results were obtained when the IVF medium included 0.5 mg P+1.0 mg PVA per milliliter with no more than 10 mM Hepes, and when 3.0 mg PVA/ml and 10 mM Hepes were present for IVC. Different concentrations of NaHCO3, up to 50 mM from 25 mM, during IVF did not alter results. Embryos produced in defined conditions yielding the best results remained viable after vitrification as evidenced by continued development in vitro for 96 h postthawing. Bovine oocytes matured in defined medium supplemented with LH were fertilized and cultured up to the blastocyst stage in chemically defined conditions that afforded results comparable to those reported earlier after inclusion of serum, BSA, and/or somatic cells.
“…Among other components investigated during in vitro maturation, fertilization, and culture (IVMFC) are sodium bicarbonate (NaHCO 3 ) [32,33]; insulin, transferrin, and selenium [34]; growth factors [35,36]; gas atmosphere [37]; glucose [38]; penicillamine (P) [39]; antioxidants [40]; and water quality [41]. However, completely defined conditions have been unavailable for definitive testing; either serum or BSA has been included for one or more of the steps composing the IVMFC procedure.…”
The objective was to establish an in vitro system in which bovine oocytes can be matured, fertilized, and cultured up to the blastocyst stage without support of serum, BSA, or somatic cells. Media consisted of modified tissue culture medium 199 (mTCM 199) with ovine LH (oLH) for maturation (IVM), experimental alterations of modified defined medium (mDM) for sperm selection and insemination (IVF), and citrate+synthetic oviductal fluid+nonessential amino acids (c-SOF+NEA) for zygote/embryo culture (IVC). Effects of heparin, BSA, polyvinyl alcohol (PVA), penicillamine (P), Hepes, and sodium bicarbonate (NaHCO3) were studied. Results included proportions of oocytes that cleaved by 48 h and that reached morulae by 120 h, blastocysts by 168 h, and expanded blastocysts by 216 h postinsemination (pi). Best results were obtained when the IVF medium included 0.5 mg P+1.0 mg PVA per milliliter with no more than 10 mM Hepes, and when 3.0 mg PVA/ml and 10 mM Hepes were present for IVC. Different concentrations of NaHCO3, up to 50 mM from 25 mM, during IVF did not alter results. Embryos produced in defined conditions yielding the best results remained viable after vitrification as evidenced by continued development in vitro for 96 h postthawing. Bovine oocytes matured in defined medium supplemented with LH were fertilized and cultured up to the blastocyst stage in chemically defined conditions that afforded results comparable to those reported earlier after inclusion of serum, BSA, and/or somatic cells.
“…Co-culture with somatic cells affects embryo development-not only secretion of growth factors but also glucose reduction caused by their metabolism [7,27]. Kim et al [17] reported that supplementation TCM199 with 5.56 mM glucose suppressed embryo development, and the same effect of glucose was reported in other culture media [18,22,30,34]. Therefore, it is considered that glucose as a component of TCM199 delays and reduces the development of embryos in vitro.…”
ABSTRACT. The present study was conducted to examine the effects of culture systems and culture media on developmental competence and freezability of bovine embryos obtained by in vitro culture of in vitro matured and fertilized (IVM-IVF) oocytes. No significant difference was observed in the proportions of oocytes developed to blastocysts, the speed at which the oocytes reached the blastocyst stage and the number of cells, when the IVM-IVF oocytes were cultured in CR1aa with or without cumulus cells. Nevertheless, more of the IVM-IVF oocytes cultured either with or without cumulus cells in CR1aa were seen to reach the blastocyst stage much sooner than those cultured with cumulus cells in TCM199 (P<0.05). The proportion of embryos developed to the blastocyst stage by day 7 in CR1aa culture was significantly higher than embryos cultured in TCM199. Viability after frozen-thawed blastocysts were obtained in vitro, was seen in a significantly higher percentage of embryos cultured in TCM199 and developed to the hatched blastocysts than in those cultured in CR1aa (P<0.05). These results indicate that CR1aa was superior to TCM199 for the potential developmental of IVM-IVF oocytes to blastocysts during in vitro culture regardless of co-culture with or without cumulus cells. But the freezability of blastocysts developed in CR1aa was inferior to those developed in TCM199. KEY WORDS: culture medium, developmental competence, freezability, in vitro culture, in vitro produced bovine embryo.J. Vet. Med. Sci. 64(10): 887-891, 2002 In vitro culture of bovine embryos derived from IVM-IVF have succeeded in producing calves [10,21]. An adequate in vitro culture system for bovine zygotes is required for large scale embryo production by IVM-IVF and genetic improvement by means of ovum pick-up and IVM-IVF. This culture system is used for the development and efficient utilization of embryonic techniques in cattle, for example for embryonic or somatic cell nuclear transfer and in transgenic animals. Bovine IVM-IVF zygotes were first cultured with cumulus cells [8,10], oviduct epithelial cells [9,21], granulosa cells [13], amnion cells [2] and buffalo rat liver cells [36] in TCM199 supplemented with bovine serum, but there are many differences between in vitro produced bovine embryos and those produced in vivo. It was reported that in vitro produced embryos have darker cytoplasm and lower density [24,25]. CR1aa medium was developed for in vitro culture of bovine IVM-IVF zygotes [15,29,30], but the optimum culture system for IVM-IVF oocytes and freezability of blastocysts cultured in CR1aa still remain to be clarified. The chilling sensitivity should be considered as a factor affecting the quality of in vitro produced bovine blastocysts.The aim of the present study was to examine the effects of culture systems and culture media on the development competence and the quality of bovine embryos obtained by in vitro culture of IVM-IVF oocytes.
MATERIALS AND METHODS
Collection of oocytes and in vitro maturation:Bovine ovaries were obtained ...
“…This shift towards glycolysis appears to be accompanied by a decreasing requirement for mitochondrial oxidative phosphorylation, as post-compaction stage embryo development appears to benefit from a reduction in oxidative phosphorylation during in vitro culture [18,19]. The culture of embryos in conditions that do not support such stage-specific metabolic preferences also provide for less than optimal development [20]. Although there is no d i r e c t e v i d e n c e t o d a t e , p e r t u r b a t i o n s i n subsequent fetal development is likely to result from metabolic conditions that cause stress to e m b r y o s a n d r e s u l t i n l e s s t h a n o p t i m a l development in vitro, as this has been reported for other species, such as the mouse [21].…”
Section: Metabolism Is a Key Regulator Of Embryo Developmentmentioning
Abstract. Cumulus-oocyte complexes (COCs) and early embryos rely on a histotrophic nutrition source for energy production and the synthesis of macromolecules. There is accumulating evidence suggesting that the balance of supply and demand for energy and other anabolic substrates during oocyte maturation and very early stages of development programmes subsequent developmental potential, and this may include subsequent fetal growth trajectory. One example is the role of glucose (Glc) during cumulus-oocyte complex maturation. Glucose is an essential nutrient for maturation, especially its role during cumulus expansion. Our laboratory has shown that during in vitro culture, too little glucose during cumulus-oocyte complex maturation affects meiotic competence. We have focussed on glucose (Glc) metabolism through the hexosamine biosynthesis pathway (HBP) during COC maturation in vitro. The HBP in somatic cells is regarded as a "fuel-sensing" pathway and its interaction with cell signalling systems and transcriptional regulation is increasingly apparent. Upregulation of the HBP during oocyte maturation in vitro has negative consequences for subsequent development. Another example is the role of hypoxia (low O2) during peri-compaction development. My laboratory believes that ruminant embryos during compaction, blastulation and subsequent development in the uterine cavity lack a key hypoxia responsive element. Because of this, hypoxia is important for normal development in ruminants but perturbs further development in rodents. The implication of these examples to the fundamental concept of peri-conception nutritional programming of development are discussed. Key words: Glucose, Oxygen, Oocyte, Embryo, Development (J. Reprod. Dev. 52: [169][170][171][172][173][174][175] 2006) he ruminant cumulus-oocyte complex from antral follicles and early embryo are both surrounded by fluids, from which they obtain their nutrients and release the products of metabolic activity. Cumulus-oocyte complexes are largely surrounded by the antral follicular fluid, apart from the "stalk" of granulosa cells which connect the mural granulosa to the cumulus-oocyte complex (see Fig. 1). Immediately following ovulation, much of the cumulus vestment surrounding the ruminant embryo is lost [1,2], such that fertilization and early development occur whilst surrounded by oviduct and uterine fluids. Indeed, the ruminant e m b r y o i s c o m p l e t e l y d e p e n d e n t o n t h i s histotrophic nutrition source for an extended p e r i o d o f d e v e l o p m e n t r e l a t i v e t o o t h e r m a m m a l i a n s p e ci es , a s i t f or m s n o m a j o r attachment to the uterine lumen until well after initial organogenesis occurs, in the cow this does not occur until at least Day 19 of development [3].Both in vitro maturation of the cumulus-oocyte complex and embryo culture of ruminant embryos are associated with impaired developmental outcomes. This is mostly in the form of a significant reduction in survival following transfer [4][5][6] and/or ...
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.