In vitro development and embryo transfer of sperm-injected oocytes derived from pregnant mares

Meintjes, M.; Graft, K.J.; Paccamonti, D.; Eilts, B.E.; Cochran, R.; Sullivan, Michael; Fall, H.; Godke, R.A.

doi:10.1016/0093-691x(96)84777-7

Cited by 20 publications

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“…In particular, the current techniques for in vitro oocyte maturation, conventional IVF, and culture of zygotes to the blastocyst stage are inadequate. For example, although only two foals have been produced via conventional IVF [1,6], the adoption of ICSI [3,7] has, by circumventing the need for sperm to penetrate the zona pellucida (ZP), resulted in the birth of several foals [3,[8][9][10][11]. Still, most ICSI pregnancies have resulted from transferring immediate postcleavage zygotes to the oviduct of recipient mares [3,8,9], whereas only a few have been produced via IVC to the blastocyst stage and transfer to the uterus [4,10,11].…”

Section: Introductionmentioning

confidence: 99%

Effects of In Vitro Production on Horse Embryo Morphology, Cytoskeletal Characteristics, and Blastocyst Capsule Formation

Tremoleda

Stout

Lagutina

et al. 2003

Biology of Reproduction

123

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Blastocyst formation rates during horse embryo in vitro production (IVP) are disappointing, and embryos that blastulate in culture fail to produce the characteristic and vital glycoprotein capsule. The aim of this study was to evaluate the impact of IVP on horse embryo development and capsule formation. IVP embryos were produced by intracytoplasmic sperm injection of in vitro matured oocytes and either culture in synthetic oviduct fluid (SOF) or temporary transfer to the oviduct of a ewe. Control embryos were flushed from the uterus of mares 6-9 days after ovulation. Embryo morphology was evaluated with light microscopy, and multiphoton scanning confocal microscopy was used to examine the distribution of microfilaments (AlexaFluor-Phalloidin stained) and the rate of apoptosis (cells with fragmented or terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling-positive nuclei). To examine the influence of culture on capsule formation, conceptuses were stained with a monoclonal antibody specific for capsular glycoproteins (OC-1). The blastocyst rate was higher for zygotes transferred to a sheep's oviduct (16%) than for those cultured in SOF (6.3%). Day 7 IVP embryos were small and compact with relatively few cells, little or no blastocoele, and an indistinct inner cell mass. IVP embryos had high percentages of apoptotic cells (10% versus 0.3% for in vivo embryos) and irregularly distributed microfilaments. Although they secreted capsular glycoproteins, the latter did not form a normal capsule but instead permeated into the zona pellucida or remained in patches on the trophectodermal surface. These results demonstrate that the initial layer of capsule is composed of OC-1-reactive glycoproteins and that embryo development ex vivo is retarded and aberrant, with capsule formation failing as a result of failed glycoprotein aggregation.

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Section: Introductionmentioning

confidence: 99%

Effects of In Vitro Production on Horse Embryo Morphology, Cytoskeletal Characteristics, and Blastocyst Capsule Formation

Tremoleda

Stout

Lagutina

et al. 2003

Biology of Reproduction

123

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“…For instance, in vitro maturation (IVM) of isolated oocytes has yielded high rates of oocytes reaching the metaphase II stage (reviewed by Dell'Aquila et al [3]); the meiotic competence of the cumulus-oocyte complex after IVM Accepted April 12, 1999. Received October 22, 1998. 1 Part of the work has been presented at the following meetings: 14th Scientific Meeting of the European Society of Embryo Transfer (AETE), Venice, Italy, 11-12 September 1998 has been investigated [4,5]; acrosome reaction following in vitro sperm capacitation treatments has been observed [6][7][8]; sperm-zona binding has been reported [7,9,10]; the oocyte's ability to sustain pronuclear formation and early embryonic development has been observed, mainly following the application of ICSI [3,[11][12][13][14][15][16][17][18]; high fertilization rates have been achieved after oviductal transfer of oocytes that had been cultured in vitro after beginning maturation in vivo [19][20][21]; and also high rates of fertilization have been observed if the zona pellucida (ZP) has been dissected [22] or drilled [23]. However, recent studies on the application of conventional IVF procedures still show low fertilization rates of in vitro-matured oocytes after various sperm treatments [8,24,25].…”

Section: Introductionmentioning

confidence: 99%

Effects of Fetuin on Zona Pellucida Hardening and Fertilizability of Equine Oocytes Matured In Vitro1

Dell’Aquila

Felici

Massari

et al. 1999

Biology of Reproduction

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In vitro fertilization (IVF) has had poor success in the horse, a situation related to low rates of sperm penetration through the zona pellucida (ZP). Zona pellucida hardening (ZPH) is seen in mouse and rat oocytes cultured in serum-free medium. The hardened ZP is refractory to sperm penetration. Fetuin, a component of fetal calf serum, inhibits ZPH and allows normal fertilization rates in oocytes cultured in the absence of serum. We evaluated whether fetuin is present in horse serum and follicular fluid (FF) and whether fetuin could inhibit ZPH in equine oocytes matured in vitro, thus increasing sperm penetration during IVF. The presence of fetuin in equine serum and FF was confirmed by immunoblotting. Oocytes submitted to in vitro maturation (IVM) in medium containing fetuin were used for ZPH assay or IVF. Intracytoplasmic sperm injection (ICSI) was carried out as a control procedure. The presence of fetuin during IVM did not affect the rate of maturation to metaphase II. Maturation of oocytes in the presence of fetuin reduced ZPH in a dose-dependent manner. After both IVF and ICSI, there was no significant difference in oocyte fertilization between fetuin-treated and untreated oocytes. The fertilization rate was significantly higher after ICSI than after IVF, both in fetuin-treated and in untreated oocytes. In conclusion, fetuin reduced ZPH in equine oocytes but did not improve sperm penetration during IVF. This implies that, in the horse, "spontaneous" ZPH is unlikely to be the major factor responsible for inhibiting sperm penetration in vitro.

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“…There are a few reports on ICSI in pig [20,24], horse [25][26][27][28][29][30] and sheep [20,31,32]. The same as ICSI in cattle, cleavage rates were low in these domestic animals.…”

Section: Problems For Practical Use Of Icsi In Pig Horse and Sheepmentioning

confidence: 99%

“…However, the cleavage rate of oocytes after ICSI was very low (12 percent, 61/ 507) [4], the production rate of transferable embryos was low for practical use [4,[12][13][14][15][16][17][18][19][20][21][22][23][24]. In other domestic animals (pig, horse and sheep), the cleavage rate of ICSI oocytes was also low [20,[24][25][26][27][28][29][30][31][32].There are many excellent reviews on the current status and future of ICSI in humans [33], mice [34] and domestic animals including cattle [35][36][37][38][39][40], and this pa--Reviewper reviews the problems with and improvements to ICSI for practical use in domestic animals, especially in cattle. …”

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confidence: 99%

Intracytoplasmic Sperm Injection (ICSI) in Cattle and Other Domestic Animals: Problems and Improvements in Practical Use.

Horiuchi

Numabe

1999

J.Mamm.Ova.Res.

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Intracytoplasmic sperm injection (ICSI) is the [8][9][10].In human ICSI, high fertilization and pregnancy rates were achieved when a motile spermatozoon was injected after immobilization [6]. Dramatic advances in the treatment of severe male-factor infertility have been achieved by using ICSI, and at present, the technique of ICSI is being widely used in clinical applications for human infertility.In cattle, the first ICSI calves were born in 1990 in Japan [4]. These results showed that bovine embryos produced by ICSI with a dead (killed) bull spermatozoon could develop into live calves. The normality of the calves (growth, behavior and reproduction) was also reported after five years [11]. However, the cleavage rate of oocytes after ICSI was very low (12 percent, 61/ 507) [4], the production rate of transferable embryos was low for practical use [4,[12][13][14][15][16][17][18][19][20][21][22][23][24]. In other domestic animals (pig, horse and sheep), the cleavage rate of ICSI oocytes was also low [20,[24][25][26][27][28][29][30][31][32].There are many excellent reviews on the current status and future of ICSI in humans [33], mice [34] and domestic animals including cattle [35][36][37][38][39][40], and this pa--Reviewper reviews the problems with and improvements to ICSI for practical use in domestic animals, especially in cattle.

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In vitro development and embryo transfer of sperm-injected oocytes derived from pregnant mares

Cited by 20 publications

References 0 publications

Effects of In Vitro Production on Horse Embryo Morphology, Cytoskeletal Characteristics, and Blastocyst Capsule Formation

Effects of In Vitro Production on Horse Embryo Morphology, Cytoskeletal Characteristics, and Blastocyst Capsule Formation

Effects of Fetuin on Zona Pellucida Hardening and Fertilizability of Equine Oocytes Matured In Vitro1

Intracytoplasmic Sperm Injection (ICSI) in Cattle and Other Domestic Animals: Problems and Improvements in Practical Use.

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