Culture of Two- and Four-Cell Rabbit Embryos to the Expanding Blastocyst Stage in Synthetic Media

Kane, MT; Foote, R.H.

doi:10.3181/00379727-133-34595

Cited by 140 publications

(53 citation statements)

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“…Using the data of Daniel (1964) for the number of cells per embryo, the present experiments indicated that a decline in the rate of protein synthesis on a per embryonic cell basis continues to occur after blastocyst formation. These differences in results may reflect the need to supplement culture medium with amino acids and serum albumin to obtain development of rabbit embryos beyond the morula stage (Kane and Foote 1970a;1970b;, and suggest that the rate of formation of amino acids from glucose is not sufficient to meet all the requirements for the continuing development of the rabbit embryo beyond the early blastocyst stage. The accumulation of label in the lipid fraction of the embryos probably arises from the diversion of glucose carbon into acetate and glycerol.…”

Section: Discussionmentioning

confidence: 98%

The in Vitro Metabolism of [U-14C]Glucose by the Preimplantation Rabbit Embryo

Quinn¹,

Wales²

1973

Aust. Jnl. Of Bio. Sci.

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The incorporation of glucose carbon into various components of embryos and culture medium was studied throughout the preimplantation period of development in the rabbit embryo.During the first 3 days after ovulation, embryos were able to develop for 24 hr in a simple chemically defined medium containing glucose and serum albumin. After blastocyst formation, this medium could maintain normal morphological appearance of day 4, day 5, and day 6 blastocysts for at least 3-4 hr.The uptake of glucose carbon by the embryo increased as development progressed, with substantial increases in the incorporation of glucose occurring after blastocyst formation. A considerable amount of the label incorporated was present in the embryos as the parent substrate, indicating that the uptake of glucose was greater than its conversion to other metabolites. Nevertheless, throughout development glucose carbon was incorporated into a wide variety of the carbon pools of the embryo, such as acidic and basic acid-soluble compounds, proteins, and lipids. Lactate, pyruvate, and acetate also accumulated in the culture medium. The rate of protein synthesis declined in relation to the increase in cell number during development and may reflect the requirements of the rabbit embryo for exogenous protein and amino acids for blastocyst expansion.A considerable amount of the incorporation of substrate carbon into day 6 blastocysts was due to the accumulation of lactate, pyruvate, acetate, amino acids, and the parent substrate within the blastocoel fluid. The higher concentration of metabolites, such as carboxylic acids and amino acids, in the blastocoel fluid than in the incubation medium indicated that there was some active mechanism for retaining these compounds within the blastocyst. The blastocyst tissue also contained labelled metabolites such as lactate, citrate, and malate as well as the amino acids glutamate, alanine, and aspartate. The accumulation of glucose carbon in these metabolites indicates that glucose is metabolized in the rabbit embryo by the operation of the Embden-Meyerhof pathway and tricarboxylic acid cycle.

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Section: Discussionmentioning

confidence: 98%

The in Vitro Metabolism of [U-14C]Glucose by the Preimplantation Rabbit Embryo

Quinn¹,

Wales²

1973

Aust. Jnl. Of Bio. Sci.

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“…In the present study, cell proliferation in Day 4 blastocysts cultured for 48 h was only insignificantly higher than in Day 4 controls, unless uterine constituents were provided in vitro (Table 3). After early growth, Day 5 blastocysts started to degenerate as early as 24 h in vitro ( (Kane & Foote, 1970). Criteria other than morphological staging (Fischer & Beier, 1986) and sequential estimates with various stages, however, seem to be more appropriate for comparing different media.…”

Section: Discussionmentioning

confidence: 99%

“…The basic culture media used were BSM II (Kane, 1969;Maurer, 1978) and (Ham, 1963 (1980) have shown that the length of the S-phase of the cell cycle for cultured 3-day-old preimplantation mouse embryos was about 7-8 h, and G¡ and G2 + M 2-3 h each. The labelling of more than 50% of embryoblast and trophoblast cells after a 4-h incubation period was substantiated in rabbit embryos by autoradiography (unpub¬ lished observation).…”

Section: Methodsmentioning

confidence: 99%

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Development retardation in cultured preimplantation rabbit embryos

Fischer¹

1987

Reproduction

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“…no. A 7511), 108 mM-NaCl, 4-78 mM-KCl, l-71-mM-CaCl2.2H20, 119 mM-KH2P04, 119 mM-MgS04.7H20, 25 mMNaHC03, 0-5 mM-sodium pyruvate, 1 mM-glucose, 100 units penicillin G/ml, 50 pg streptomycin sulphate/ml and the amino acids, vitamins and trace elements of Ham's FIO medium (Ham, 1963;Kane & Foote, 1970). The charcoal-treated BSA was added to the basic medium (1) as a type of BSA from which most small molecules have been removed, (2) because it is possible that rabbit blastocyst development requires protein in the medium for physicochemical reasons, and (3) because it has been suggested that BSA can protect embryos against toxic substances and preliminary work showed that high levels of the low molecular weight extract of BSA were toxic to the embryos.…”

Section: Methodsmentioning

confidence: 99%

Culture of Two- and Four-Cell Rabbit Embryos to the Expanding Blastocyst Stage in Synthetic Media

Cited by 140 publications

References 1 publication

The in Vitro Metabolism of [U-14C]Glucose by the Preimplantation Rabbit Embryo

The in Vitro Metabolism of [U-14C]Glucose by the Preimplantation Rabbit Embryo

Development retardation in cultured preimplantation rabbit embryos

A low molecular weight extract of bovine serum albumin stimulates rabbit blastocyst cell division and expansion in vitro

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