Morphology of Day 7 Bovine Demi-Embryos during in vitro Reorganization

Albihn, Ann; Rodríguez‐Martínez, H.; Gustafsson, Helena

doi:10.1159/000146918

Cited by 12 publications

(16 citation statements)

References 6 publications

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“…The in vivo-derived blastocysts have numerous large microvilli projecting from the trophoblast cells into the perivitelline space. Similar findings were observed by other investigators (Linares and Ploen 1981;Mohr and Trounson 1981;Albihn et al 1990;. On the other hand, the in vitrodeveloped blastocysts have poorly developed microvilli.…”

Section: Discussionsupporting

confidence: 91%

“…The observable differences in ultrastructural features between embryos obtained non-surgically from superovulated cows and the embryos developed from IVM-IVF oocytes in a serum-supplemented medium were as follows: (1) Previous studies reported that lipid droplets were present in large numbers in the bovine oocytes and early cleavage stages of embryos but were significantly reduced in blastocysts (Brackett et al 1980;Mohr and Trounson 1981;Albihn et al 1990;Shamsuddin et al 1993b). Plante and King (1994) also compared the amount of lipid droplets in the cytoplasm of in vivo-derived and the in vitro-developed bovine embryos.…”

Section: Discussionmentioning

confidence: 99%

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Fine structure of bovine morulae and blastocysts in vivo and in vitro

Abé¹,

Otoi²,

Tachikawa³

et al. 1999

Anatomy and Embryology

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The ultrastructure of bovine morulae and blastocysts developed from in vitro-matured and -fertilized oocytes in a serum-supplemented medium was compared with that of morulae and blastocysts collected non-surgically from superovulated cows. In the in vivo-derived morulae, two characteristic cells types could be identified by the electron-density of their cytoplasm and by their ultrastructural features. One type appeared light in color with low electron-dense cytoplasm. These cells were located in the peripheral layer of the cluster of blastomeres, possessed numerous cellular organelles such as mitochondria and Golgi apparatus and had microvilli projecting into the perivitelline space. The other cell type was distinguished by cytoplasm that stained more densely than that of the lighter-appearing cells. The darker-appearing cells generally possessed fewer organelles than the lighter cells, but many lysosome-like structures were present in the cytoplasm. The in vitro-developed morulae also contained two types of cells similar to those observed in the in vivo morulae. However, most of the in vitro-developed cells possessed numerous lipid droplets and contained fewer lysosome-like structures than the cells of the in vivo-derived morulae. The blastocysts, both in vivo and in vitro, showed a clear differentiation of trophoblast cells and inner cell mass (ICM)-cells. In the in vivo-derived blastocyst, the apical membrane of trophoblast cells was covered with large, numerous microvilli and well-developed junctional complexes were observed. Lipid droplets were present in the cytoplasm of trophoblast and ICM-cells but were not abundant. In vitro-developed blastocysts showed less well-developed junctional complexes between trophoblast cells, less well-developed apical microvilli on the trophoblast cells, and contained large numbers of lipid droplets. This accumulation of lipid droplets was higher in the trophoblast cells than in the ICM-cells. The zonae pellucidae of in vitro-developed embryos were thinner than that of the in vivo-derived embryos. This study demonstrates conspicuous differences in the ultrastructural features between the in vivo-derived and in vitro-developed embryos, suggesting that the ultrastructure may reflect the various physiological anomalies observed in previous studies.

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Section: Discussionsupporting

confidence: 91%

Section: Discussionmentioning

confidence: 99%

Fine structure of bovine morulae and blastocysts in vivo and in vitro

Abé¹,

Otoi²,

Tachikawa³

et al. 1999

Anatomy and Embryology

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“…It has also been reported that bovine embryos bisected and cultured for 4 to 6 h survived cryopreservation [6]. Within 30 min after bisection, the bovine demi-embryos were found to have restored their prebisection morphology such as cell polarization, cell-to-cell contact between the trophoblastic and/or inner cell mass cells and the presence of mitosis [17]. Therefore, since the membrane and structure of the cell of biopsied embryos damaged by surgery may recover during these culture periods, the tolerance of the embryos may be improved.…”

Section: Discussionmentioning

confidence: 79%

Effect of Time Interval between Biopsy and Vitrification on Survival of In Vitro-Produced Bovine Blastocysts.

Ito¹,

Sekimoto

Hirabayashi³

et al. 1999

Journal of Reproduction and Development

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Abstract. The aim of this study was to improve the cryosurvival of bovine blastocysts following biopsy. In vitro-derived bovine blastocysts were biopsied with a metal blade, then cultured for 0 to 20 h in m-SOF and then vitrified in the presence of ethylene glycol and dimethyl sulfoxide. The post-warm survival rates after 24 h of in vitro culturing of the biopsied embryos were significantly higher at 1.25, 2.5, 5 and 10 h than that of 0 and 20 h (69, 73, 78 and 64% vs 34 and 27%, p<0.05, respectively). The survival rate of embryos vitrified at 2.5 and 5 h after biopsy was not different from that of the non-vitrified control group (73 and 78% vs 96%, p=0.08 vs 0.07, respectively). These results indicate that 1.25 to 10 h culture between biopsy and vitrification improves the viability of bovine blastocysts produced in vitro.

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“…Normal day 7 bovine blastocysts have numerous large microvilli projecting from the trophoblast cells into the perivitelline space (LINARES and PLOEN, 1981;MOHR and TROUNSON, 1981 ;ALBIHN et al, 1990). These microvilli are essential for the fluid transportation activity of an embryo ( B E~E R I D G E and FLECHON, 1988).…”

Section: Flechonmentioning

confidence: 99%

“…Cytoplasmic vesicles and lipid droplets, which are present in large amounts in the bovine oocytes and early cleavage stages of embryos, get significantly reduced in day 7 blastocysts ( B R A C K E~ et al, 1980;MOHR and TROUNSON, 1981;ALBIHN et al, 1990;. Embryos with large amounts of vesicles and high lipid content showed more sensitivity to cryopreservation procedures (MOHR and TROUNSON, 1981;LEIBO and LOSKUTOFF, 1993).…”

Section: Flechonmentioning

confidence: 99%

Fine Structure of Bovine Blastocysts Developed Either in Serum‐Free Medium or in Conventional Co‐Culture With Oviduct Epithelial Cells

Shamsuddin

Rodriguez‐Martinez

1994

Journal of Veterinary Medicine Series A

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Summary The ultrastructure of day 7 bovine blastocysts developed in vitro in either of two culture systems was compared with that of morphologically normal blastocytes collected non‐surgically from superovulated cows on day 7 (Day 0 = day of insemination). The in vitro‐embryos were obtained after culture of in vitro‐matured and ‐fertilized oocytes either in a serum‐free, cell‐free medium (SFM, i.e. TCM 199 supplemented with BSA (10 mg/ml), insulin (5 μg/ml), transferrin (5 μg/ml) and selenium (5ng/ml) or in a serum‐supplemented medium (TCM 199 and 10% (v/v) oestrous cow serum) together with bovine oviduct epithelial cells (BOEC). Five of the 8 blastocysts developed in SFM fulfilled the criteria set for normal morphology of the in vivo‐developed blastocytes. In contrast, 6 out of 8 blastocysts developed in co‐culture with BOEC were classified as morphologically deviated, and only 2 reached the criteria for morphological normality. In vitro‐developed blastocysts with deviated morphology showed a higher degree of cytoplasmic vacuolation, short, less developed cell‐to‐cell contacts between trophoblast as well as between inner cell mass (icm)‐cells, less developed apical microvilli on the trophoblast and wide inter‐cellular spaces. Additionally, numerous cytoplasmic vesicles, phagosomes, lipid droplets and hooded mitochondria were commonly present, both in trophoblast and in icm‐cells. The results indicate that a high proportion of blastocysts developed in co‐culture with BOEC were morphologically deviated compared to those cultured in medium where serum and somatic cells were replaced by BSA, insulin, transferrin and selenium.

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Morphology of Day 7 Bovine Demi-Embryos during in vitro Reorganization

Cited by 12 publications

References 6 publications

Fine structure of bovine morulae and blastocysts in vivo and in vitro

Fine structure of bovine morulae and blastocysts in vivo and in vitro

Effect of Time Interval between Biopsy and Vitrification on Survival of In Vitro-Produced Bovine Blastocysts.

Fine Structure of Bovine Blastocysts Developed Either in Serum‐Free Medium or in Conventional Co‐Culture With Oviduct Epithelial Cells

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