The effects of amino acids, cumulus cells, and bovine serum albumin on in vitro fertilization and first cleavage of hamster eggs

The effects of glutamine, hypotaurine, taurine and premixed solutions of essential amino acids (EAA) and non-essential amino acids (NEAA) on in vitro development of porcine zygotes were evaluated. The effects of refreshing the medium and replacing polyvinyl alcohol (PVA) with bovine serum albumin (BSA) on embryonic development were also investigated. Porcine zygotes produced by in vitro maturation (IVM) and in vitro fertilisation (IVF) were cultured in porcine zygote medium (PZM), as the basal culture medium, for 5 days after IVF. The total number of cells in blastocysts was significantly increased by the addition of 2 mm glutamine to PZM, as was blastocyst yields after supplementation with 0.25 to 4 mm glutamine. Addition of 1.25 to 10 mm hypotaurine to PZM significantly increased blastocyst yields. Addition of 5 mm taurine to PZM significantly increased blastocyst yield, whereas taurine had no effect on blastocyst yield in cultures already containing 5 mm hypotaurine. Adding 1× EAA significantly increased the rate of blastocyst formation compared with no or 2× EAA, whereas 2× NEAA significantly increased the total cell numbers in blastocysts compared with no NEAA. Refreshing the medium at Day 3 had no effect on blastocyst yields, whereas medium change significantly reduced the total cell numbers in blastocysts. Adjusting the amino acid concentrations of a chemically defined medium can improve the developmental competence of porcine embryo.

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Stimulation of rat ovarian ornithine decarboxylase in vitro by hCG, amino acids and bovine serum albumin

Acta Endocrinologica ◽

10.1530/acta.0.0940571 ◽

1980 ◽

Vol 94 (4) ◽

pp. 571-576 ◽

Cited By ~ 3

Author(s):

Kirsti Käpyaho

Keyword(s):

Amino Acids ◽

Bovine Serum Albumin ◽

Cyclic Amp ◽

Serum Albumin ◽

Ornithine Decarboxylase ◽

Bovine Serum ◽

Defined Medium ◽

Decarboxylase Activity ◽

Ornithine Decarboxylase Activity

Abstract. Rat ovarian ornithine decarboxylase activity could be stimulated in vitro by a variety of factors, which apparently have different modes of action. Ovarian cells prepared from pre-pubertal rats by collagenase dispersion exhibited a low but detectable ornithine decarboxylase activity after a 6-h incubation in a defined medium. The enzyme activity was markedly enhanced in vitro by hCG, which also produced increased accumulation of cyclic AMP and stimulated the secretion of progesterone. In addition to the gonadotrophin, ovarian ornithine decarboxylase activity was strikingly stimulated by some non-essential amino acids, and especially by bovine serum albumin. While markedly enhancing ornithine decarboxylase activity, none of the latter additions increased the accumulation of cyclic AMP or enhanced the secretion of progesterone. Bovine serum albumin enhanced powerfully ornithine decarboxylase activity in vitro at very small concentrations (from 0.75 μm). The half-life of the enzyme remained unchanged (26—28 min) upon stimulation indicating that the stimulation mechanism did not involve any stabilization of the enzyme.

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Attempts at in vitro fertilization and culture of in vitro matured oocytes in sei (Balaenoptera borealis) and Bryde's (B. edeni) whales

Zygote ◽

10.1017/s0967199408004887 ◽

2009 ◽

Vol 17 (1) ◽

pp. 19-28 ◽

Cited By ~ 6

Author(s):

M. M. U. Bhuiyan ◽

Y. Suzuki ◽

H. Watanabe ◽

K. Matsuoka ◽

Y. Fujise ◽

...

Keyword(s):

In Vitro Fertilization ◽

Bovine Serum Albumin ◽

Embryo Culture ◽

Culture Medium ◽

Cumulus Cells ◽

Protein Supplementation ◽

Vitro Fertilization ◽

Embryo Culture Medium ◽

Morula Stage

SummaryThe cumulus–oocyte–complexes (COCs) recovery rates with respect to reproductive status per sei (Balaenoptera borealis) and Bryde's (B. edeni) whales were determined in Experiment 1. The number of COCs recovered ranged from 16.0 to 30.6 and from 6.7 to 26.8 per sei and Bryde's whales, respectively. The effects of COCs grades and protein supplementation in embryo culture medium on development of in vitro fertilized (IVF) embryos were evaluated in sei and Bryde's whales in Experiment 2. The COCs were classified into either Grade A (COCs with five or more layers of compact cumulus cells) or Grade B (COCs with less than five layers of compact or expanded cumulus cells) before being cultured for IVM. The cleavage (12.0 to 19.5%), 4-cell (8.0 to 12.0%) and 8-cell (4.0 to 8.0%) formation rates in sei whales did not vary significantly between embryos derived from either grade A or B oocytes and between embryos cultured in either fetal whale serum (FWS)- or bovine serum albumin (BSA)-supplemented medium. The cleavage (4.0 to 14.8%), 4-cell (0.0 to 7.5%) and 8-cell (0.0 to 2.6%) formation rates in Bryde's whales did not vary significantly between embryos derived from either grade A or B oocytes and between embryos cultured in either FWS- or BSA-supplemented medium. The grade B oocytes cultured in FWS-supplemented medium developed to morula stage (1.1%) in sei whales. In conclusion, the present study indicates that IVF in sei whales is possible to achieve cleaved embryos developing to morula stage. This is the first in vitro embryo production attempt in sei and Bryde's whales.

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A Brief Incubation of Cumulus-Enclosed Mouse Eggs in a Calcium-Free Medium Containing a High Concentration of Calcium-Chelator Markedly Improves Preimplantation Development

International Journal of Environmental Research and Public Health ◽

10.3390/ijerph17103505 ◽

2020 ◽

Vol 17 (10) ◽

pp. 3505

Author(s):

Valeria Merico ◽

Silvia Garagna ◽

Maurizio Zuccotti

Keyword(s):

In Vitro Fertilization ◽

Mammalian Species ◽

Developmental Rate ◽

Cumulus Cells ◽

Fertilization Rate ◽

Preimplantation Development ◽

High Concentration ◽

Vitro Fertilization ◽

Positive Effects

The presence of cumulus cells (CCs) surrounding ovulated eggs is beneficial to in vitro fertilization and preimplantation development outcomes in several mammalian species. In the mouse, this contribution has a negligible effect on the fertilization rate; however, it is not yet clear whether it has positive effects on preimplantation development. Here, we compared the rates of in vitro fertilization and preimplantation development of ovulated B6C3F1 CC-enclosed vs. CC-free eggs, the latter obtained either after a 5 min treatment in M2 medium containing hyaluronidase or after 5–25 min in M2 medium supplemented with 34.2 mM EDTA (M2-EDTA). We found that, although the maintenance of CCs around ovulated eggs does not increment their developmental rate to blastocyst, the quality of the latter is significantly enhanced. Most importantly, for the first time, we describe a further quantitative and qualitative improvement, on preimplantation development, when CC-enclosed eggs are isolated from the oviducts in M2-EDTA and left in this medium for a total of 5 min prior to sperm insemination. Altogether, our results establish an important advancement in mouse IVF procedures that would be now interesting to test on other mammalian species.

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A Precipitin-like Reaction of the Hemolymph of the Lobster Homarus americanus

Journal of the Fisheries Research Board of Canada ◽

10.1139/f69-127 ◽

1969 ◽

Vol 26 (5) ◽

pp. 1392-1397 ◽

Cited By ~ 10

Author(s):

James E. Stewart ◽

Diane M. Foley

Keyword(s):

Bovine Serum Albumin ◽

Serum Albumin ◽

Bovine Serum ◽

Serum Proteins ◽

Homarus Americanus ◽

Test Period ◽

Fluorescent Material

The levels of fluorescent material in the hemolymph of lobsters injected with serum proteins from lobster hemolymph labelled with fluorescein remained relatively constant over a 6-day test period; the levels in lobsters injected with bovine serum albumin labelled with fluorescein declined rapidly. A precipitin-like reaction was observed when lobster hemolymph serum was titrated with bovine serum albumin in vitro.

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