Sucrose-induced shrinkage of in vitro produced bovine morulae: Effect on viability, morphology and ease of evaluation

Soom, Ann Van; Ysebaert, M.T.; Medts, A. Vanhoucke-De; Velde, A. Van de; Merton, S.; Delval, A.; Langendonckt, Anne Van; Donnay, Isabelle; Vanroose, G.; Bols, P.E.J.; Kruif, Aart de

doi:10.1016/s0093-691x(96)00285-3

Cited by 13 publications

(13 citation statements)

References 27 publications

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“…Agreement among evaluators is reduced when determinating stage of development of compact morulae produced in vitro [12,13]. Furthermore, pregnancy rates following transfer of compact morulae produced in vitro [14] were lower than those obtained after transfer of compact morulae produced in vivo [15].…”

Section: Discussionmentioning

confidence: 96%

“…Compared with embryos produced in vivo, embryos produced in vitro have fewer total blastomeres [7], demonstrate incomplete compaction [8,9] and cell-to-cell coupling [10], and have a greater variance in morphological quality and developmental rate [11]. Differences in morphology may contribute to the decrease in agreement among evaluators when determining stage of development for compact morulae produced in vitro [12,13]. Furthermore, it is interesting to note that pregnancy rates after transfer of morula-stage bovine embryos produced in vitro [14] were lower than pregnancy rates after the transfer of morulae produced in vivo [15].…”

Section: Introductionmentioning

confidence: 99%

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Ultrastructural Morphometry of Bovine Compact Morulae Produced In Vivo or In Vitro1

Crosier

Farin

Dykstra

et al. 2000

Biology of Reproduction

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The objective of this study was to compare the ultrastructure of bovine compact morulae produced in vivo or in vitro using morphometric analysis. Compact morulae produced in vivo were obtained from superovulated Holstein cows. Compact morulae produced in vitro were obtained from cumulus-oocyte complexes aspirated from ovaries of Holstein cows. The complexes were matured and fertilized in vitro. At 20 h postinsemination (hpi), zygotes were distributed into 1 of 3 culture media: 1) IVPS (in vitro produced with serum): TCM-199 + 10% estrous cow serum (ECS); 2) IVPSR (in vitro produced with serum restriction): TCM-199 + 1% BSA until 72 hpi followed by TCM-199 + 10% ECS from 72 to 144 hpi; 3) mSOF (modified synthetic oviductal fluid): SOF + 0.6% BSA. At 144 hpi, five grade 1 compact morulae from each of the four treatments were prepared for transmission electron microscopy. The volume density occupied by cellular components was determined by the point-count method using a sampling of seven to nine random micrographs from each compact morula. The volume density of lipid was greater (P < 0.05) in compact morulae from IVPS, IVPSR, and mSOF treatments compared with those produced in vivo. There was a reduced proportional volume of total mitochondria in compact morulae from the IVPS treatment compared with those produced in vivo (P < 0.05). For compact morulae from the IVPS culture treatment, the volume density of vacuoles was greater than that for compact morulae produced in vivo (P < 0.05). The cytoplasmic-to-nuclear ratio for compact morulae from the IVPS treatment was increased (P < 0.05) compared with the ratio for those produced in vivo. In conclusion, compact morulae produced in vitro differed ultrastructurally from those produced in vivo. Compact morulae produced in IVPS culture medium possessed the greatest deviations in cellular ultrastructure.

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Section: Discussionmentioning

confidence: 96%

Section: Introductionmentioning

confidence: 99%

Ultrastructural Morphometry of Bovine Compact Morulae Produced In Vivo or In Vitro1

Crosier

Farin

Dykstra

et al. 2000

Biology of Reproduction

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“…Embryos that developed into blastocysts were categorized into one of three groups, good, fair or poor, by microscopic observation of the blastocyst morphology (Van Soom et al 1996). Embryos that developed into blastocysts were categorized into one of three groups, good, fair or poor, by microscopic observation of the blastocyst morphology (Van Soom et al 1996).…”

Section: Ivm Ivf and Ivc Proceduresmentioning

confidence: 99%

“…Development to the four-cell embryo stage and blastocyst formation was evaluated after 24 h and 7 days of culture, respectively. Embryos that developed into blastocysts were categorized into one of three groups, good, fair or poor, by microscopic observation of the blastocyst morphology (Van Soom et al 1996). Embryos were classified as 'good' if their appearance was as expected for the respective stage of development with few, if any, extraneous degenerated cells.…”

Section: Ivm Ivf and Ivc Proceduresmentioning

confidence: 99%

In vitro development of OPU‐derived bovine embryos cultured either individually or in groups with the silk protein sericin and the viability of frozen‐thawed embryos after transfer

Isobe

Ikebata²,

Kim

et al. 2014

Animal Science Journal

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The optimization of single-embryo culture conditions is very important, particularly in the in vitro production of bovine embryos using the ovum pick-up (OPU) procedure. The purpose of this study was to examine the development of embryos derived from oocytes obtained by OPU that were cultured either individually or in groups in medium supplemented with or without sericin and to investigate the viability of the frozen-thawed embryos after a direct transfer. When two-cell-stage embryos were cultured either individually or in groups for 7 days in CR1aa medium supplemented with or without 0.5% sericin, the rates of development to blastocysts and freezable blastocysts were significantly lower for the embryos cultured individually without sericin than for the embryos cultured in groups with or without sericin. Moreover, the rate of development to freezable blastocysts of the embryos cultured individually with sericin was significantly higher than that of the embryos cultured without sericin. When the frozen-thawed embryos were transferred directly to recipients, the rates of pregnancy, abortion, stillbirth and normal calving in the recipients were similar among the groups, irrespective of the culture conditions and sericin supplementation. Our findings indicate that supplementation with sericin during embryo culture improves the quality of the embryos cultured individually but not the viability of the frozen-thawed embryos after transfer to recipients.

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“…Mechanical isolation of embryos from the ZP was performed as follows. Groups of five to 10 embryos were placed in 25 µL of 0.3 m sucrose (Van Soom et al 1996) covered with mineral oil until shrinkage of embryos occurred. Thereafter, most of the 0.3 m sucrose was removed such that the weight of the mineral oil aided in maintaining the embryos in place.…”

Section: Alkaline Phosphatase Activity After Enzymatic or Mechanical mentioning

confidence: 99%

Alkaline phosphatase activity in bovine oocytes and preimplantation embryos as affected by removal of the zona pellucida and culture medium constituents

Edwards¹,

Powell²,

Rexroad³

2003

Reprod. Fertil. Dev.

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The aims of the present study were: (1) to characterize alkaline phosphatase (AP) activity in bovine oocytes and embryos with intact or removed zona pellucida (ZP); and (2) to evaluate the effect of culture medium constituents on AP activity. Alkaline phosphatase activity in non-matured and matured oocytes was most evident nearest the plasma membrane and perivitelline space. In more than 90% of two- to 16-cell embryos, AP activity was observed in the perivitelline space and at blastomere contacts. In blastocysts, AP activity was localized to the trophectoderm. Only after immunodissection was AP activity detected in the inner cell mass. Removal of the ZP by pronase or mechanical means reduced AP activity. Alkaline phosphatase activity was detected in evacuated ZP after mechanical removal. Specific constituents comprising the embryo culture medium altered AP activity. Alkaline phosphatase activity was reduced in eight- to 16-cell embryos and evacuated ZP cultured in CR1aa + 0.4% bovine serum albumin compared with embryos cultured in CR1aa alone or embryos co-cultured on a monolayer of Buffalo rat liver cells in the presence of 10% fetal bovine serum. The presence of AP activity at blastomere contacts and in evacuated ZP limits its usefulness as a marker for the differentiation of embryonic cells comprising the early cleavage-stage embryo.

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Sucrose-induced shrinkage of in vitro produced bovine morulae: Effect on viability, morphology and ease of evaluation

Cited by 13 publications

References 27 publications

Ultrastructural Morphometry of Bovine Compact Morulae Produced In Vivo or In Vitro1

Ultrastructural Morphometry of Bovine Compact Morulae Produced In Vivo or In Vitro1

In vitro development of OPU‐derived bovine embryos cultured either individually or in groups with the silk protein sericin and the viability of frozen‐thawed embryos after transfer

Alkaline phosphatase activity in bovine oocytes and preimplantation embryos as affected by removal of the zona pellucida and culture medium constituents

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