Stimulation of early embryonic development in the sheep by co-culture with oviduct epithelial cells
TL;DR: It is concluded that during the first 3 days after fertilization cleavage will progress at a normal rate on different feeder-layers but oviduct cells appear to be required for the acquisition of full embryonic viability.
Abstract: To examine the effects of somatic cell support on the cleavage and viability of fertilized sheep eggs, 434 pronucleate eggs were co-cultured for 3 or 6 days on oviduct cells or fibroblasts and 77 eggs were cultured in medium alone. During the first 3 days in culture 95% of the single-celled eggs cleaved regularly to non-compacted morulae on either of the feeder-layers but only 13% underwent similar regular cleavage in medium alone. Despite the identical cleavage rates in the co-culture groups, only 33% of embryos grown on fibroblasts as compared with 80% of embryos grown on oviduct cells were fully viable as judged by their ability to develop normally after transfer to recipient animals. The viability of embryos in the oviduct group was equal to that obtained after the direct transfer of morulae from donor to recipient sheep. After 6 days in culture 42% of embryos co-cultured with oviduct cells developed into expanded blastocysts as compared with only 4.5% cultured on fibroblasts. In both co-culture groups virtually all the remaining embryos blocked during the 4th cleavage. When transferred, 30% of blastocysts grown from the pronucleate stage on oviduct cells were viable. We conclude that: (1) during the first 3 days after fertilization cleavage will progress at a normal rate on different feeder-layers but oviduct cells appear to be required for the acquisition of full embryonic viability.(ABSTRACT TRUNCATED AT 250 WORDS)
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TL;DR: La presente revue considere les donnees recentes concernant les produits impliques dans l'activation du genome de l'embryon chez plusieurs especes.
Abstract: La presente revue considere les donnees recentes concernant les produits impliques dans l'activation du genome de l'embryon chez plusieurs especes
884 citations
Cites background from "Stimulation of early embryonic deve..."
...The sheep embryo also exhibits a block at the eight- to 16cell stage when one-cell embryos are cultured in vitro in medium alone, but a very high proportion of viable morulae and blastocysts can be obtained when fertilized eggs are cocultured with oviduct cells (Gandolfi and Moor, 1987)....
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TL;DR: Clear evidence is presented that specific growth factors of embryonic and/or reproductive tract origin participate in preimplantation embryo development and blastocyst functions in an autocrine/paracrine manner.
Abstract: We have established a model that shows cooperative interaction among preimplantation embryos and the role of growth factors on their development and growth. Two-cell mouse embryos cultured singly in 25-microliters microdrops had inferior development to blastocysts and lower cell numbers per blastocyst compared with those cultured in groups of 5 or 10. The inferior development of singly cultured embryos was markedly improved by addition of epidermal growth factor (EGF) or transforming growth factor alpha or beta 1 (TGF-alpha or TGF-beta 1) to the culture medium. The stage of embryonic development, primarily affected by these treatments, was between eight-cell/morula and blastocyst. Furthermore, blastocysts developed from eight-cell embryos cultured in groups or singly in the presence of EGF showed a higher incidence of zona hatching compared with those cultured singly in the absence of EGF. Detection of EGF receptors on the embryonic cell surface at eight-cell/morula and blastocyst stages suggests beneficial effects of EGF or TGF-alpha on preimplantation embryo development and blastocyst functions. Insulin-like growth factor I (IGF-I) had no influence on embryo development. To further document the cooperative interactions among embryos, the volume of the culture medium was doubled to 50 microliters. This increase in culture volume was even more detrimental to the development of singly cultured embryos. However, this detrimental effect was significantly reversed by EGF and reversed even more markedly by a combination of EGF and TGF-beta 1 but not by TGF-beta 1 alone. Although TGF-beta 1 plus IGF-I caused a modest improvement of embryo development, the response was not as great as shown by EGF alone. Furthermore, IGF-I had no additive effect on EGF-induced embryonic development. The study presents clear evidence that specific growth factors of embryonic and/or reproductive tract origin participate in preimplantation embryo development and blastocyst functions in an autocrine/paracrine manner.
677 citations
TL;DR: Investigation of sheep zygote development of amino acids, ammonium, vitamins, and culture of embryos in groups in Synthetic Oviduct Fluid medium supplemented with BSA found indirect evidence that ruminant embryos utilize amino acids to a greater extent than do rodent embryos.
Abstract: The aim of this study was to develop a serum-free culture system that could support high levels of cleavage and blastocyst formation from sheep zygotes developed in vitro. To this end, we investigated the effects on sheep zygote development of amino acids, ammonium, vitamins, and culture of embryos in groups in Synthetic Oviduct Fluid (SOF) medium supplemented with BSA (32 mg/ml). The inclusion of amino acids in the culture medium had no effect on the percentage of embryos arrested at the 8-16-cell stage when embryos were cultured singly in the same drop of medium for 6 days (43% in SOF; 41% in SOF+amino acids). However, in medium containing all Eagle's amino acids, replacing the culture medium every 48 h to alleviate ammonium toxicity significantly decreased the number of arrested embryos (6%; p < 0.05) and significantly increased blastocyst cell number (52 cells in SOF; 105 cells in SOF+amino acids; p < 0.01) and the number of embryos developing to the blastocyst stage (29% in SOF; 67% in SOF+amino acids; p < 0.05). When the medium was renewed every 48 h, nonessential amino acids and glutamine also significantly decreased the number of arrested embryos (p < 0.05). Culturing embryos singly or in groups in SOF medium with all Eagle's amino acids that was renewed every 48 h resulted in significant increases in blastocyst hatching and mean cell number (47%, 31%, and 79%; 105, 136, and 173 cells for embryos cultured singly, in groups of 2, and in groups of 4, respectively). After culture in groups of 4, blastocyst cell numbers were equivalent to in vivo-developed controls (160 cells) and significantly greater than those developed in serum (103 cells; p < 0.01). Analysis of blastocyst metabolism, expressed on a per-cell basis, revealed that amino acids did not affect either glucose uptake or lactate production, whereas the addition of amino acids and vitamins resulted in a significant increase in both parameters (p < 0.01). A similar response was observed in serum-derived blastocysts. Ammonium production by sheep blastocysts after culture in the presence of amino acids was significantly greater than that produced by mouse blastocysts, indirect evidence that ruminant embryos utilize amino acids to a greater extent than do rodent embryos.(ABSTRACT TRUNCATED AT 400 WORDS)
558 citations
TL;DR: Five-8-cell embryos from superovulated cattle were co-cultured with oviducal tissue suspended in Ham's F10 + 10% fetal calf serum or in F10FCS alone and embryos obtained from in-vitro maturation and fertilization were used to compare development between co-culture and medium conditioned by ovidUCal tissue.
Abstract: In Exp. 1, 5-8-cell embryos from superovulated cattle were co-cultured with oviducal tissue suspended in Ham's F10 + 10% fetal calf serum (F10FCS) or in F10FCS alone. After 4 days, the proportion of embryos developing into compact morulae or blastocysts was greater (P less than 0.005) in co-culture (38/82; 46%) than in F10FCS (1/27; 4%). In Exp. 2, a solution of collagenase, trypsin, DNAse and EDTA was used to disperse oviducal tissue, which was then cultured in TCM199 + 10% fetal calf serum (M199FCS) to obtain monolayers. Embryos (1-8 cells) were then co-cultured with monolayers or in M199FCS alone. The proportion of embryos developing into compact morulae and blastocysts after 4-5 days was higher (P less than 0.005) in co-culture (15/34; 43%) than in M199FCS (1/37; 3%); mean numbers of cells/embryo were also higher (P less than 0.001) (27.70; range 2-82 in co-culture; 8.83; range 2-18 in M199FCS). In Exp. 3, embryos obtained from in-vitro maturation and fertilization were used to compare development between co-culture and medium conditioned by oviducal tissue. Initial cleavage rate (no. embryos greater than 1 cell/total) was 76% (611/807) and did not differ among treatments. After 5 days, the proportion cleaving to greater than 16 cells was higher (P less than 0.005) in co-culture (71/203; 35%) and conditioned medium (48/205; 23%) compared to M199FCS (14/203; 7%). Similarly, the proportion developing into compact morulae and blastocysts was greater (P less than 0.005) in co-culture (44/203; 22%) and conditioned medium (46/205; 22%) than in M199FCS (7/203; 3%).(ABSTRACT TRUNCATED AT 250 WORDS)
482 citations
Cites background from "Stimulation of early embryonic deve..."
...Among the large domestic species, early sheep embryos have been cultured past the 8-16-cell block to the blastocyst stage on oviducal cell monolayers (Gandolfi & Moor, 1987)....
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...More recently, oviducal cell monolayers have been shown to enhance the development of sheep embryos in vitro (Gandolfi & Moor, 1987; Rexroad & Powell, 1988)....
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TL;DR: The oviduct is a muscular tube with a mucous lining that differs considerably between species, notably in the degree of coiling, and in the best-characterized species in this respect are the cow and the rabbit.
Abstract: Gabriele Fallopius (1523-1562) is credited with the first correct anatomical description of the oviduct, which he termed the "tuba uteri" after its resemblance to a brass musical instrument. Nowadays the terms 'oviduct', 'Fallopian tube', 'uterine tube' and 'tube' tend to be used interchangeably. In this review, the term 'oviduct' will be used to refer to the mammal generally, the term 'Fallopian tube' to the human exclusively. The oviduct is a muscular tube with a mucous lining. However, its gross anatomy differs considerably between species, notably in the degree of coiling. It is usually divided into 4 regions; the infundibulum, fringed by fimbriae and opening via the tuba1 ostium into the peritoneal cavity, the ampulla, the isthmus and the uterotubal junction. Some authors term the infundibulum and fimbriae the preampulla; the uterotubal junction is also known as the intramural or interstitial portion. For most of its length, the oviduct has two muscle coats, an external longitudinal and an internal circular coat. The mucosa is thrown into folds, also known as 'rugae' or 'plicae'. The proportions of muscle and mucosa vary along the length of the oviduct. The best-characterized species in this respect are the cow and the rabbit (El-Banna & Hafez, 1970; Leese, 1983), in both of which there is a gradual decline in the proportion of mucosa, the degree of mucosal folding, and the surface area of the lumen, and an increase in the proportion of muscle, as one proceeds from the infundibulum to the uterotubal junction.
408 citations
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TL;DR: This communication describes the successful culture of one-cell to eight-cell sheep ova and one- cell and eight- cell cattle ova to the morula and blastocyst stages and reports a high embryo survival after transfer of cultured Ova to recipient animals.
Abstract: Fertilized sheep and cattle ova have not been reported to develop readily during culture in vitro. Up to 60% of sheep morulae develop normally during culture (Moor & Cragle, 1971) but earlier cleavage stages undergo limited development (Hancock, 1963; Kraemer, 1966; Tervit & McDonald, 1969; Moore, 1970) and it has been suggested that there is a block to development in vitro at the eightto twelve-cell stage (Wintenberger, Dauzier & Thibault, 1953). Only the early cleavage stages of cattle ova have been cultured and these have not been reported to develop beyond the twenty-four-cell stage in vitro (Thibault, 1966; Brinster, 1968; Sreenan, 1968; Sreenan, Scanlon & Gordon, 1968). This communication describes the successful culture of one-cell to eight-cell sheep ova and one-cell and eight-cell cattle ova to the morula and blastocyst stages and reports a high embryo survival after transfer of cultured ova to recipient animals.
1,009 citations
"Stimulation of early embryonic deve..." refers background in this paper
...The presence of somatic cells increases the rate of cleavage and development of sheep zygotes in culture to levels well in advance of those achieved previously ( Tervit et al., 1972; Tervit & Rowson, 1974; Wright & Bondioli, 1981)....
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TL;DR: An examination of the spermatozoa and oocyte incubation media showed no media or protein supplement to be superior in promoting in vitro fertilization in cattle, sheep or swine.
Abstract: Summary Various aspects of the major components of in vitro fertilization and embryo culture in domestic farm animals are discussed. An examination of the spermatozoa and oocyte incubation media showed no media or protein supplement to be superior in promoting in vitro fertilization in cattle, sheep or swine. Generally, ovulated oocytes or activated follicular oocytes were penetrated by sperm more frequently than were immature oocytes. Spermatozoa that were incubated in vivo or in the oviducts of a different species generally achieved higher oocyte penetration than did spermatozoa incubated in vitro. In most in vitro fertilization studies, an in vivo component was introduced, generally oocyte or sperm maturation, which served to confound analysis of the in vitro results. Furthermore, the relatively low level of success and high incidence of chromosomal abnormalities with in vitro fertilization in other species require that careful and complete studies analyzing the components of in vitro fertilization in domestic farm animals be conducted. The culture of embryos from domestic farm animals is detailed, with emphasis on in vitro conditions and rate of success. As with laboratory animals, bovine, ovine, porcine and caprine embyos of fewer than eight cells are more difficult to culture to the blastocyst
264 citations
"Stimulation of early embryonic deve..." refers background in this paper
...The presence of somatic cells increases the rate of cleavage and development of sheep zygotes in culture to levels well in advance of those achieved previously (Tervit et al., 1972; Tervit & Rowson, 1974; Wright & Bondioli, 1981 )....
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...This is particularly true of domestic animals for which only a small minority of long-term cultured embryos cleave regularly and even this minority almost invariably fail to develop further when transferred to recipient animals (reviewed by Wright & Bondioli, 1981 )....
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TL;DR: Embryos which had spent 3 days in an advanced recipient were transferred to a recipient synchronous with the original donor, and embryos first transferred on Day 6 were markedly stimulated and less able to implant in the second recipient.
Abstract: Sheep embryos which were transferred to recipients in oestrus 3 days before or 3 days after the donors were unable to implant even when a synchronously transferred embryo was developing successfully in the same uterus. Embryos which had spent 3 days in an advanced recipient were transferred to a recipient synchronous with the original donor. Embryos first transferred on Day 3 were slightly accelerated in their development, but retained the ability to implant normally in the 2nd recipient. By contrast, embryos first transferred on Day 6 were markedly stimulated and less able to implant in the second recipient.
157 citations
"Stimulation of early embryonic deve..." refers background in this paper
...Equally it could be due to the requirements for subtle uterine signals analogous to those known to influence embryonic development in sheep between Days 9 and 12 ( Wilmut & Sales, 1981; Godkin et al., 1984)....
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TL;DR: It is shown that a high percentage of zygotes cleave to two cell stages and then cease development when cultured in Waymouth medium supplemented with ATP, deoxynucleosides and a feeder layer of irradiated HeLa cells, suggesting that the initial development of the mouse zygote is dependent on a supply of exogenous factors, additional to those required by later stages.
Abstract: TECHNIQUES have been perfected for the culture in vitro of late two cell mouse ova into blastocysts in simple chemically defined conditions1,2. Attempts to obtain development from earlier stages have failed, unless the ova were cultured in organ cultures of the fallopian tube3,4. These results suggested that the fallopian tube provides some critical contribution necessary for the development of the mouse zygote into the late two cell stage. Nevertheless, it has been a common experience that when the mouse zygote is cultivated independently of the fallopian tube a few cleave to the two cell stage and then cease development5. It has now been shown6, however, that a high percentage of zygotes cleave to two cell stages and then cease development when cultured in Waymouth7 medium supplemented with ATP, deoxynucleosides and a feeder layer of irradiated HeLa cells. These experiments suggest that the initial development of the mouse zygote is dependent on a supply of exogenous factors, additional to those required by later stages.
105 citations
"Stimulation of early embryonic deve..." refers background in this paper
...Further work on the role of the maternal environment in supporting passage through the transition phase is required, especially since oviduct expiants can support the development of mouse embryos across the 2-cell block ( Whittingham & Biggers, 1967 )....
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