Effect of medium renewal during culture in two different culture systems on development to blastocysts from in vitro produced early bovine embryos.

1996 ◽  
Vol 74 (11) ◽  
pp. 2752 ◽  
Author(s):  
Y Fukui ◽  
E S Lee ◽  
N Araki
Keyword(s):  
Bovine Embryos ◽  
Culture Systems

Evaluation of different culture systems on the in vitro production of bovine embryos

2005 ◽  
Vol 63 (4) ◽  
pp. 1131-1141 ◽  
Author(s):  
Daniela Costa Pereira ◽  
Margot Alves Nunes Dode ◽  
Rodolfo Rumpf
Keyword(s):  
Bovine Embryos ◽  
In Vitro Production ◽  
Culture Systems ◽  
Vitro Production

166 EFFECT OF INDIVIDUAL CULTURE SYSTEM ON IN VITRO DEVELOPMENT OF IN VITRO-MATURED - IN VITRO-FERTILIZED BOVINE EMBRYOS

2012 ◽  
Vol 24 (1) ◽  
pp. 195
Author(s):  
R. Nishii ◽  
K. Imai ◽  
H. Koyama ◽  
O. Dochi
Keyword(s):  
Culture System ◽  
Calf Serum ◽  
Control Culture ◽  
Total Cell ◽  
Bovine Embryo ◽  
Bovine Embryos ◽  
Culture Systems ◽  
Individual Culture ◽  
Single Culture

An individual in vitro culture system for bovine embryo needs to be developed for the study of embryo developmental competence. The objective of this study was to examine the effect of individual culture systems on the development of in vitro-matured–in vitro-fertilized bovine embryos. Two individual culture systems were compared. Cumulus–oocyte complexes (COC) were collected by aspiration of ovarian follicles (diameter, 2 to 5 mm) obtained from a local abattoir. The COC were matured in TCM-199 supplemented with 5% calf serum and 0.02 AU mL–1 of FSH. Groups of 20 COC were incubated in 100-μL droplets of IVM media at 38.5°C under an atmosphere of 5% CO2 for 20 h. After 18 h of gamete co-culture (3 × 106 sperm mL–1), the presumptive zygotes were cultured in CR1aa medium supplemented with 5% calf serum at 38.5°C under an atmosphere of 5% CO2, 5% O2 and 90% N2 for 9 days (fertilization = Day 0). The presumptive zygotes were randomly assigned to 1 of the following 3 treatments: single culture (1 zygote was cultured in a 5-μL droplet), well-of-the-well (WOW; Sugimura et al. 2010 Biol. Reprod. 83, 970–978) culture (25 zygotes were cultured individually in each 125-μL droplet) and control culture (25 zygotes were cultured in a 125-μL droplet). Embryo development was evaluated for cleavage and blastocyst rates, on Day 2 and Day 7 to 9 after IVF, respectively. The rates of development up to the blastocyst stage and total cell number in the blastocysts, determined by an air-drying method, were investigated. The cleavage and blastocyst rates were analysed by the chi-square test and the total cell numbers were analysed by ANOVA. The cleavage rates were significantly higher in the control and WOW groups than in the single-culture group (P < 0.01) and the blastocyst rates were significantly lower in the single-culture group than in the control culture group (P < 0.05; Table 1). The total cell numbers (mean ± s.d.) of blastocysts did not significantly differ between the single culture (154.6 ± 21.8), control culture (155.2 ± 22.5) and WOW culture (159.8 ± 27.0) groups. These results indicate that although the blastocyst rate was lower in the single-culture system than in the WOW or group culture system, in vitro-matured–in vitro-fertilized bovine embryos can be cultured using the single-culture system. Moreover, the quality of blastocysts developed by the single-culture system is the same as that of blastocysts developed using the other 2 culture systems. Table 1.Effect of different culture methods for bovine embryo development

Use of conditioned medium for IVM-IVF bovine embryos in-vitro culture systems

1993 ◽  
Vol 39 (1) ◽  
pp. 236 ◽  
Author(s):  
W.G. van Inzen ◽  
Th.A.M. Kruip ◽  
S.M. Weima
Keyword(s):  
In Vitro Culture ◽  
Conditioned Medium ◽  
Bovine Embryos ◽  
Culture Systems

Development of bovine embryos cultured in CR1aa and IVD101 media using different oxygen tensions and culture systems

2010 ◽  
Vol 58 (4) ◽  
pp. 465-474 ◽  
Author(s):  
Tamás Somfai ◽  
Yasushi Inaba ◽  
Yoshio Aikawa ◽  
Masaki Ohtake ◽  
Shuji Kobayashi ◽  
...  
Keyword(s):  
Inner Cell Mass ◽  
Culture Media ◽  
Cell Mass ◽  
Calf Serum ◽  
Bovine Embryos ◽  
Blastocyst Formation ◽  
Cell Numbers ◽  
Culture Systems ◽  
Significant Difference

The aim of the present study was to optimise the culture conditions for the in vitro production of bovine embryos. The development of in vitro fertilised bovine oocytes in CR1aa supplemented with 5% calf serum and IVD101 culture media were compared using traditional microdrops and Well of the Well (WOW) culture systems either under 5% or 20% oxygen tension. After 7 days of culture, a significantly higher blastocyst formation rate was obtained for embryos cultured in CR1aa medium compared to those cultured in IVD101, irrespective of O 2 tensions and culture systems. The blastocyst formation in IVD101 was suppressed under 20% O 2 compared to 5% O 2 . Despite their similar total cell numbers, higher rates of inner cell mass (ICM) cells were observed in blastocysts developed in IVD101 medium than in those developed in CR1aa, irrespective of O 2 tensions. There was no significant difference in blastocyst formation, total, ICM and trophectoderm (TE) cell numbers between embryos obtained by microdrop and WOW culture systems irrespective of the culture media and O 2 tensions used. In conclusion, CR1aa resulted in higher blastocyst formation rates irrespective of O 2 tension, whereas IVD101 supported blastocyst formation only under low O 2 levels but enhanced the proliferation of ICM cells.

Development and de novo protein synthetic activity of bovine embryos produced in vitro in different culture systems

2001 ◽  
Vol 55 (2) ◽  
pp. 593-606 ◽  
Author(s):  
M. Kuran ◽  
J.J. Robinson ◽  
M.E. Staines ◽  
T.G. McEvoy
Keyword(s):  
De Novo ◽  
Synthetic Activity ◽  
Bovine Embryos ◽  
Culture Systems

scholarly journals Changes in the relative abundance of mRNA transcripts for insulin-like growth factor (IGF-I and IGF-II) ligands and their receptors (IGF-IR/IGF-IIR) in preimplantation bovine embryos derived from different in vitro systems

Reproduction ◽  
2001 ◽  
pp. 601-610 ◽  
Author(s):  
MA Yaseen ◽  
C Wrenzycki ◽  
D Herrmann ◽  
JW Carnwath ◽  
H Niemann
Keyword(s):  
Growth Factor ◽  
Polyvinyl Alcohol ◽  
Blastocyst Stage ◽  
Bovine Embryo ◽  
Bovine Embryos ◽  
Cell Stage ◽  
Culture Systems ◽  
Igf I ◽  
Oviduct Fluid

The aim of this study was to determine the relative abundance of mRNAs for the insulin-like growth factor I (IGF-I) and IGF-II ligands, and for the IGF receptors (IGF-IR and IGF-IIR) in in vitro preimplantation bovine embryos from the oocyte to the hatched blastocyst stage using two different culture systems: TCM-199 supplemented with oestrous cow serum, or synthetic oviduct fluid supplemented with polyvinyl alcohol. Development to the two- to four-cell stage and blastocyst stage was significantly higher (P < or = 0.05) in embryos cultured in TCM supplemented with oestrous cow serum than in those cultured in synthetic oviduct fluid supplemented with polyvinyl alcohol (61 and 25% versus 55 and 17%, respectively). A semi-quantitative RT-PCR assay did not detect IGF-I transcripts at any stage of preimplantation bovine development, including the hatched blastocyst stage. In both culture systems, IGF-IR, IGF-II and IGF-IIR were expressed throughout preimplantation development up to the hatched blastocyst stage in a varying pattern. The expression patterns of IGF-IR, IGF-II and IGF-IIR in embryos generated in the two culture systems were not significantly different, except at the expanded blastocyst stage, at which significantly higher amounts of IGF-IIR were observed in the TCM system than in the synthetic oviduct fluid system. These results indicate that transcripts of IGF-IR and IGF-IIR follow the standard pattern in which maternal stores of mRNA in the oocyte are slowly depleted up to the 16-cell stage and then re-established at the onset of embryonic expression of these genes. The lack of detectable IGF-I transcripts in the bovine embryo indicates a predominantly paracrine mode of action. The bovine embryo is capable of producing IGF-II, IGF-IIR and IGF-IR in large amounts, particularly after hatching, which may be important for the formation of the filamentous conceptus. Results indicate an autocrine mechanism for IGF-II and modulation of IGF family expression by culture conditions.

279 IN VITRO DEVELOPMENT IN THREE CULTURE SYSTEMS OF BOVINE OOCYTES FERTILIZED WITH SEX-SORTED SPERM

2011 ◽  
Vol 23 (1) ◽  
pp. 237
Author(s):  
B. Trigal ◽  
E. Gomez ◽  
M. Muñoz ◽  
J.N. Caamaño ◽  
S. Carrocera ◽  
...  
Keyword(s):  
Culture System ◽  
Final Analysis ◽  
Vero Cells ◽  
Bovine Embryos ◽  
Vitro Fertilization ◽  
In Vitro Development ◽  
Culture Systems ◽  
Cell Counts ◽  
Vitro Development

Following in vitro fertilization and culture, the timing of both embryonic cleavage and appearance of blastocysts has been reported to be altered when using sorted spermatozoa (Lu et al. 1999 Theriogenology 52, 1393–1405). To optimize an in vitro culture system for bovine embryos produced with sexed sperm, sorted and unsorted (control) semen samples of 5 bulls (semen samples provided by XY Inc., Fort Collins, CO, USA) were separated using a Percoll gradient followed by a washing step with Fert-TALP medium (Trigal et al. 2010 Reprod. Domest. Anim. 45, 83). The obtained pellet was used for fertilizing in vitro matured cumulus–oocyte complexes from slaughterhouse ovaries. Zygotes were cultured in 1) Vero cells monolayer + B2 + 10% FCS; 2) SOFaaci + 10% FCS; or 3) SOFaaci + 6 g L–1 BSA. A total of 1980 oocytes were in vitro matured and fertilized. Development was recorded on Days 3 and Days 6 to 8, and analysed by ANOVA and Duncan test. Bull, replicate, and culture effects were nonsignificant, and consequently were not considered in the final analysis. Data are percentages of development related to the zygotes cultured, and are expressed as LSM ± SE (see Table 1). The use of sexed sperm significantly reduced development rates compared with unsorted sperm. Thus, sorted sperm showed reduced 5- to 8-cell, Day-6 morulae, and Day-8 blastocyst rates over unsorted sperm within the 3 culture systems analysed. Culture in SOF + BSA was the least efficient embryo culture system in terms of Day-7 blastocyst rates, both with unsorted and sorted sperm. Day-7 blastocyst rates produced with unsorted sperm after culture in Vero cells or SOF + FCS were significantly higher than those obtained after culture in SOF + BSA. Day-7 blastocyst rates obtained with sorted sperm and cultured in Vero cells or SOF + FCS were comparable to those obtained with unsorted sperm in SOF + BSA. No differences were detected between male- and female-sorted sperm (data not shown). Sorted sperm is an effective tool for producing sex-known embryos. Quality assessment (differential cell counts and cryosurvival) of embryos produced with sorted sperm under different conditions are being undertaken. Table 1.In vitro development of bovine embryos fertilized with either untreated or sorted sperm and cultured in Vero cells, SOF + FCS, or SOF + BSA Supported by INIA RTA2008.#0082. M. Muñoz, B. Trigal and D. Martin are sponsored by RYC08-03454, Cajastur and PTA2007-#0268-I. We acknowledge Sexing Technologies for collaboration.

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