Co-culture of day-5 to day-7 equine embryos in medium with oviductal tissue

Freeman, D.A.; Butler, James E.; Weber, J.A.; Geary, R.T.; Woods, G.L.

doi:10.1016/0093-691x(91)90347-g

Cited by 15 publications

(5 citation statements)

References 17 publications

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“…Hatching, associated with failure of both zona dissolution and capsule formation, has been noted previously in equine embryos recovered on Day 5 ex vivo and cultured to the blastocyst stage in vitro [31]. Freeman et al [32] also reported hatching of in vitro-cultured embryos collected ex vivo between Day 5 and Day 7, but noted that almost half of the cultured embryos experienced near-normal thinning of approximately half of the zona, and expanded within this area. These authors did not report whether the thinning was seen more commonly in embryos collected on the later days (that had been longer in the uterus).…”

Section: Discussionmentioning

confidence: 86%

Blastocyst Formation Rates In Vivo and In Vitro of In Vitro-Matured Equine Oocytes Fertilized by Intracytoplasmic Sperm Injection1

Choi¹,

Roasa²,

Love³

et al. 2004

Biology of Reproduction

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This study was conducted to evaluate in vivo and in vitro development of in vitro-matured equine oocytes fertilized by intracytoplasmic sperm injection. Oocytes were collected from slaughterhouse-derived ovaries, matured in vitro, and injected with frozen-thawed stallion sperm. In vivo development was assessed after transfer of injected oocytes to the oviducts of recipient mares. Mares were killed 7.5-8.5 days after transfer and the uterus and oviducts flushed for embryo recovery. Of 132 injected oocytes transferred, 69 (52%) were recovered; of these, 25 (36%) were blastocysts with a blastocoele and capsule. In vitro development was assessed in three culture systems. Culture of zygotes in modified Chatot, Ziomek, Bavister medium with BSA containing either 5.5 mM glucose for 7.5 days or 0.55 mM glucose for 3 days, followed by 3 mM glucose for 2 days, then 4.3 mM glucose for 2.5 days, did not result in blastocyst formation. Culture of zygotes in Dulbecco modified Eagle medium (DMEM)/F-12 with 10% fetal bovine serum with and without coculture with equine oviductal epithelial explants yielded 16% and 15% blastocyst development, respectively. Development to blastocyst was significantly lower in G1.3/2.3/BSA than in DMEM/F-12/BSA or in either medium with 10% added serum (2% vs. 18%, 18% or 20%; P < 0.05), suggesting that requirements for equine embryo development differ from those for other species. These results indicate that in vitro-matured equine oocytes are sufficiently competent to form 36% blastocysts in an optimal environment (in vivo). While we identified an in vitro culture system that provided repeatable blastocyst development without coculture, this yielded only half the rate of development achieved in vivo.

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Section: Discussionmentioning

confidence: 86%

Blastocyst Formation Rates In Vivo and In Vitro of In Vitro-Matured Equine Oocytes Fertilized by Intracytoplasmic Sperm Injection1

Choi¹,

Roasa²,

Love³

et al. 2004

Biology of Reproduction

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“…A combination of growth factors might be required for the proliferation of these pluripotent cells, as has been demonstrated in the case of cultures of mouse primordial germ cells [19]. Among studies of domestic animals, investigations of the development in vitro of equine embryos at the preimplantation stage have been quite limited [20–22]. Isolation of equine ES cells would put horses in the front lines of animal biotechnology and the modification of animal genomes.…”

Section: Introductionmentioning

confidence: 99%

Isolation of embryonic stem‐like cells from equine blastocysts and their differentiation in vitro¹

et al. 2002

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Embryonic stem (ES) cells are pluripotent cells with the potential capacity to generate any type of cell. We describe here the isolation of pluripotent ES-like cells from equine blastocysts that have been frozen and thawed. Our two lines of ESlike cells (E-1 and E-2) appear to maintain a normal diploid karyotype inde¢nitely in culture in vitro and to express markers that are characteristic of ES cells from mice, namely, alkaline phosphatase, stage-speci¢c embryonic antigen-1, STAT-3 and Oct 4. After culture of equine ES-like cells in vitro for more than 17 passages, some ES-like cells di¡erentiated to neural precursor cells in the presence of basic ¢broblast growth factor (bFGF), epidermal growth factor and platelet-derived growth factor. We also developed a protocol that resulted in the di¡er-entiation of ES-like cells in vitro to hematopoietic and endothelial cell lineages in response to bFGF, stem cell factor and oncostatin M. Our observations set the stage for future developments that may allow the use of equine ES-like cells for the treatment of neurological and hematopoietic disorders.

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“…However, in the present study, when expanding embryos were used for in-vitro culture, the maximum diameter was 1720, 2075 and 4500 pn in completely hatched embryos. Signs of degeneration were reported in embryos cultured for only 3 days in Ham's F12:DMEM (1 : I , v/v) and less frequently during coculture with oviductal tissue (Freeman et a/. 1991a).…”

Section: Discussionmentioning

confidence: 99%

“…1989): the presence or absence of a capsule before in-vitro culture may influence the manner of hatching. A third pattern of hatching, shedding gradually through a large tear in the ZP, was reported for Day 5-7 embryos, although it was not noted whether the manner of hatching was related to embryonic stage at the beginning of invitro culture (Freeman et a/. 1991a) or whether the capsule was present.…”

Section: Introductionmentioning

confidence: 98%

The hatching process of equine embryos in vitro

Hochi

Braun

Oguri

1993

Equine Veterinary Journal

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Summary Blastocysts (5 unexpanded, 5 expanding, 2 hatched) were recovered non‐surgically between 144 and 192 h after detection of ovulation. Unexpanded and expanding embryos were cultured (TCM199 + 10% foetal bovine serum) for 10 days. Mean diameter was 168.0 μm for unexpanded and 251.0 μm for expanding blastocysts, respectively. Unexpanded embryos started to hatch from the zona pellucida (ZP) when their diameter reached 405.4 μm (embryonic age 240.0 h). Four embryos squeezed through a small opening in the ZP whereas 1 embryo gradually tore open the ZP. Three embryos completed the hatching process within 96–120 h. In contrast, expanding embryos started to hatch at a mean diameter of 499.8 μm (embryonic age 192.0 h). Four embryos escaped from the ZP within 12 h. One embryo squeezed through the ZP but did not shed the ZP entirely during culture. In‐vitro hatched embryos continued to expand spherically until reaching a maximum mean diameter of 1141.7 μm (unexpanded embryos) and 2765.0 μm (expanding embryos), except for 1 embryo which attached to the bottom of the culture dish. In completely hatched embryos, no capsule could be detected at x200 magnification after staining. Three embryos which did not entirely hatch from the ZP until the end of culture had a capsule remaining inside the ZP. In 1 case, a thin acellular layer was detected in the shed ZP after 0.3% trypsin treatment. Both freshly recovered hatched blastocysts (diameter 640 μm, 697 μm) had a clearly visible capsule around the trophectoderm cells. Equine blastocysts not only lost their zona pellucida but also their capsule during the hatching process in vitro.

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Co-culture of day-5 to day-7 equine embryos in medium with oviductal tissue

Cited by 15 publications

References 17 publications

Blastocyst Formation Rates In Vivo and In Vitro of In Vitro-Matured Equine Oocytes Fertilized by Intracytoplasmic Sperm Injection1

Blastocyst Formation Rates In Vivo and In Vitro of In Vitro-Matured Equine Oocytes Fertilized by Intracytoplasmic Sperm Injection1

Isolation of embryonic stem‐like cells from equine blastocysts and their differentiation in vitro¹

The hatching process of equine embryos in vitro

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Co-culture of day-5 to day-7 equine embryos in medium with oviductal tissue

Cited by 15 publications

References 17 publications

Blastocyst Formation Rates In Vivo and In Vitro of In Vitro-Matured Equine Oocytes Fertilized by Intracytoplasmic Sperm Injection1

Blastocyst Formation Rates In Vivo and In Vitro of In Vitro-Matured Equine Oocytes Fertilized by Intracytoplasmic Sperm Injection1

Isolation of embryonic stem‐like cells from equine blastocysts and their differentiation in vitro1

The hatching process of equine embryos in vitro

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Isolation of embryonic stem‐like cells from equine blastocysts and their differentiation in vitro¹