Effect of Linoleic Acid-Albumin on Post-Thaw Survival of in vitro-Produced Bovine Embryos at the 16-Cell Stage.

Tominaga, Keiichiro; Hamada, Yukako; Yabuue, T; Ariyoshi, Tomoyuki

doi:10.1292/jvms.62.465

Cited by 19 publications

(7 citation statements)

References 19 publications

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“…However, blastocyst development and quality of 16-cell stage embryos frozen even in ethylene glycol was still inferior to those of fresh embryos (67.3%, 37/55). When the 16-cell stage embryos were produced in CR1aa [14] containing 0.25 mg/mL LAA for 4 days after IVF, the postthaw survival of 16-cell stage embryos produced in LAA-containing medium was higher than that of those cultured without LAA [15].…”

Section: Ethylene Glycol As a Permeable Cryoprotectant For Freezing Omentioning

confidence: 90%

Cryopreservation and Sexing of In Vivo- and In Vitro-Produced Bovine Embryos for Their Practical Use

Tominaga

2004

Journal of Reproduction and Development

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Abstract. My research awarded includes contributions to cryopreservation and sexing of bovine embryos produced in vitro and in vivo, as follows; (1) In vivo-derived morulae and blastocysts were cryopreserved in the presence of 10% glycerol, and the embryos were transferred into recipients after two-step dilution of glycerol in straw, with a practically acceptable pregnancy rate. (2) The survival rate of 16-cell stage embryos frozen in the medium with ethylene glycol was higher than that with DMSO or 1,2-propanediol. Addition of linoleic acid-albumin to culture medium enhanced the survival rate of post-thaw bovine 16-cell stage in vitro-produced (IVP) embryos. (3) Polarization of cytoplasmic lipid droplets by centrifugation of 2-cell stage embryos was found effective to increase freezing tolerance in 16-cell stage embryos developed from the centrifuged embryos, because blastomeres of 16-cell stage embryos were mostly lipid-free. (4) The usefulness of gel-loading tip (GLTip) as a container for ultra-rapid vitrification was demonstrated in IVP embryos from 2-cell to blastocyst stages, with a higher in vitro survival than the conventional two-step freezing. (5) PCR analysis for sexing of in vivo-derived Day-7 embryos indicated that male embryos developed faster and graded higher than female embryos. But such correlation between genetic sex and embryonic development was not found in IVP embryos obtained from individual cows. (6) Addition of 0.1-1.0% deproteinized hemodialysate product from calf blood to culture medium increased the producing efficiency of demi-embryos with good quality. Female embryos rather than male embryos required a longer time to repair after bisection. (7) In vivo-derived bovine embryos after biopsy for sexing by PCR analysis and subsequent vitrification using GL-Tips are available to practical use in the field. (8) Introduction of primer extension preamplification-PCR and purification of DNA product before standard sexing PCR of biopsy samples from Day 3-4 in vitro-derived embryos allowed accurate sex determination, and Day-7 blastocysts developed from Day 3-4 embryos were cryopreserved by GLTip vitrification without a loss of their viability. Thus the field application of bovine embryo transfer is in part supported by improvements of technologies in embryo cryopreservation and sex predetermination.

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Section: Ethylene Glycol As a Permeable Cryoprotectant For Freezing Omentioning

confidence: 90%

Cryopreservation and Sexing of In Vivo- and In Vitro-Produced Bovine Embryos for Their Practical Use

Tominaga

2004

Journal of Reproduction and Development

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“…(2006) found no difference in blastocyst production for cultures supplemented with amino acids and oestrous cow serum and/or bovine serum albumin (BSA), but post‐thaw survival rates were higher for medium containing amino acids and BSA (51.9% vs. 25%). Moreover, bovine embryos cultured in CR1aa without serum prior to vitrification had better survival post‐warming (Tominaga et al. 2000; Mucci et al.…”

Section: Cryotolerancementioning

confidence: 99%

Consequences of In Vitro Culture Conditions on Embryo Development and Quality

Rizos

Clemente

Bermejo-Álvarez

et al. 2008

Reprod Domestic Animals

183

124

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Despite major efforts directed at improving the yield of blastocysts from immature oocytes in vitro, the quality of such blastocysts continually lags behind that of blastocysts produced in vivo. These differences are manifested at the level of morphology, metabolism, gene expression and cryotolerance, and may have a knock-on effect further along the developmental axis. Evidence suggesting that in vitro culture conditions, while capable of producing blastocysts in relatively high numbers, are far from optimal with deficiencies being manifested in terms of abnormally large offspring. It is clear nowadays that modification of the post-fertilization culture environment in vitro can improve blastocyst quality to some extent.

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“…It is naturally found to be accumulated in the body fluids of many organisms as winter advances (23). Several workers have used this compound for effective cryopreservation of various tissues and cells like sperms (24), embryos (25), islets (26), RBCs (27), and other biological samples (28). Kapicht et al (29) reported freezing of human bone marrow and PBSCs with trehalose at vitrification concentrations without DMSO.…”

Section: Figmentioning

confidence: 99%

Cryopreservation of Human Hematopoietic Cells with Membrane Stabilizers and Bioantioxidants as Additives in the Conventional Freezing Medium

Limaye

Kale

2001

Journal of Hematotherapy & Stem Cell Research

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Cord blood (CB) and fetal liver (FL) cells are two alternative sources of human hematopoietic stem cells. Optimization of cryopreservation protocols is an important aspect in the banking of these tissues. Out of the multiple factors responsible for cryodamage of cells, membrane leakage and oxygen free-radical generation have been shown to contribute substantially toward the process. We have studied the effect of certain additives, like membrane stabilizers and bioantioxidants, to the conventional freezing medium on viability, nucleated cell recovery, and clonogenic potential of frozen cells. Our results show that trehalose, a membrane stabilizer, when used in combination with 10% dimethyl sulfoxide (DMSO) affords better cryoprotection as evidenced by significantly increased colony formation as compared to 10% DMSO alone. The cryoprotection afforded by trehalose persists at least for 1.5 years and there is no bias toward protection of a particular lineage. We also found that this increased cryoprotective effect of trehalose is seen both at -196 degrees C and -80 degrees C storage temperatures. Addition of taurine, an amino acid, another membrane stabilizer, and a natural cryoprotectant to the traditional freezing medium also results in beneficial effect. Of the three bioantioxidants tested, i.e., ascorbic acid, alpha-tocopherol acetate, and catalase, catalase shows maximum cryoprotective effect both at -196 degrees C and at -80 degrees C. Because the mode of cryoprotective action of catalase and trehalose are totally different, we tried a combination of these two compounds along with 10% DMSO. At -196 degrees C the protection afforded by the combination was significantly better than that afforded by individual components. At -80 degrees C, however, the combination did not give any added protection as compared to the individual single additives, although it was significantly better than 10% DMSO alone. These results indicate that the addition of membrane stabilizers and antioxidants to the conventional freezing medium may help to improve post thaw recovery of hematopoietic cells.

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Effect of Linoleic Acid-Albumin on Post-Thaw Survival of in vitro-Produced Bovine Embryos at the 16-Cell Stage.

Cited by 19 publications

References 19 publications

Cryopreservation and Sexing of <i>In Vivo</i>- and <i>In Vitro</i>-Produced Bovine Embryos for Their Practical Use

Cryopreservation and Sexing of <i>In Vivo</i>- and <i>In Vitro</i>-Produced Bovine Embryos for Their Practical Use

Consequences of In Vitro Culture Conditions on Embryo Development and Quality

Cryopreservation of Human Hematopoietic Cells with Membrane Stabilizers and Bioantioxidants as Additives in the Conventional Freezing Medium

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