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Atsuo Ogura - One of the best experts on this subject based on the ideXlab platform.

  • birth of offspring after transfer of mongolian gerbil meriones unguiculatus embryos cryopreserved by vitrification
    Molecular Reproduction and Development, 2005
    Co-Authors: Keiji Mochida, Junichiro Matsuda, Teruhiko Wakayama, Kaoru Takano, Yoko Noguchi, Yoshie Yamamoto, Osamu Suzuki, Atsuo Ogura
    Abstract:

    The Mongolian gerbil (Meriones unguiculatus) has been used as a laboratory species in many fields of research, including neurology, oncology, and parasitology. Although the cryopreservation of embryos has become a useful means to protect valuable genetic resources, its application to the Mongolian gerbil has not yet been reported. In this study, we investigated the in vitro and in vivo developmental competence of Mongolian gerbil embryos cryopreserved by vitrification. In vivo-fertilized embryos were vitrified on the day of collection using the ethylene glycol (EG)-based solutions EFS20 and EFS40, which contained 20% and 40% EG, respectively, in PB1 containing 30% (w/v) Ficoll 70 and 0.5 M sucrose. First, we compared one-step and two-step vitrification protocols. In the one-step method, the embryos were directly transferred into the vitrification solution (EFS40), whereas in the two-step method, the embryos were exposed serially to EFS20 and EFS40 and then vitrified. After liquefying (thawing), late two-cell embryos (collected on day 3) vitrified by the two-step method showed significantly better rates of in vitro development to the Morula stage compared to those vitrified by the one-step method (65% vs. 5%, P < 0.0001). We then examined whether the same two-step method could be applied to early two-cell embryos (collected on day 2), four-cell embryos (day 4), Morulae (day 5), and blastocysts (day 6). After liquefying, 87%-100% of the embryos were morphologically normal in all groups, and 23% and 96% developed to the compacted Morula stage from early two- and four-cell embryos, respectively. After transfer into recipient females, 3% (4/123), 1% (1/102), 5% (4/73), and 10% (15/155) developed to full-term offspring from vitrified and liquefied early two-cell embryos, late two-cell embryos, Morulae, and blastocysts, respectively. This demonstrates that Mongolian gerbil embryos can be safely cryopreserved using EG-based vitrification solutions.

  • birth of offspring after transfer of mongolian gerbil meriones unguiculatus embryos cryopreserved by vitrification
    Molecular Reproduction and Development, 2005
    Co-Authors: Keiji Mochida, Junichiro Matsuda, Teruhiko Wakayama, Kaoru Takano, Yoko Noguchi, Yoshie Yamamoto, Osamu Suzuki, Atsuo Ogura
    Abstract:

    The Mongolian gerbil (Meriones unguiculatus) has been used as a laboratory species in many fields of research, including neurology, oncology, and parasitology. Although the cryopreservation of embryos has become a useful means to protect valua- ble genetic resources, its application to the Mongolian gerbil has not yet been reported. In this study, we investigated the in vitro and in vivo developmental competence of Mongolian gerbil embryos cryopre- served by vitrification. In vivo-fertilized embryos were vitrified on the day of collection using the ethylene glycol (EG)-based solutions EFS20 and EFS40, which contained 20% and 40% EG, respectively, in PB1 containing 30% (w/v) Ficoll 70 and 0.5M sucrose. First, we compared one-step and two-step vitrification protocols. In the one-step method, the embryos were directly transferred into the vitrification solution (EFS40), whereas in the two-step method, the em- bryos were exposed serially to EFS20 and EFS40 and then vitrified. After liquefying (thawing), late two-cell embryos (collected on day 3) vitrified by the two-step method showed significantly better rates of in vitro development to the Morula stage compared to those vitrified by the one-step method (65% vs. 5%, P <0.0001). We then examined whether the same two-step method could be applied to early two-cell embryos (collected on day 2), four-cell embryos (day 4), Morulae (day 5), and blastocysts (day 6). After liquefying, 87%-100% of the embryos were morpho- logically normal in all groups, and 23% and 96% developed to the compacted Morula stage from early two- and four-cell embryos, respectively. After transfer into recipient females, 3% (4/123), 1% (1/102), 5% (4/73), and 10% (15/155) developed to full-term offspring from vitrified and liquefied early two-cell embryos, late two-cell embryos, Morulae, and blas- tocysts, respectively. This demonstrates that Mongo- lian gerbil embryos can be safely cryopreserved using EG-based vitrification solutions. Mol. Reprod. Dev. 70: 464-470, 2005. 2005 Wiley-Liss, Inc.

  • mouse spermatid nuclei can support full term development after premature chromosome condensation within mature oocytes
    Human Reproduction, 1999
    Co-Authors: Atsuo Ogura, Kimiko Inoue, Junichiro Matsuda
    Abstract:

    The nucleus of round spermatids, the earliest haploid male germ cells, can participate in the formation of normal zygotes when incorporated into activated oocytes. In this study, we injected mouse round spermatids into homologous mature oocytes that were kept arrested at metaphase II to induce premature chromosome condensation (PCC) of the spermatid nuclei. After full condensation of the spermatid chromosomes, the oocytes were activated by Sr 2+ -containing medium, into which cytochalasin B was added to prevent extrusion of the segregated female and male chromosomes as polar bodies. Out of 142 oocytes examined, 104 (73%) formed two male (pseudo)pronuclei and two female pronuclei. To restore the diploid state of these zygotes, one of the female pronuclei was removed. When cultured in vitro for 72 hours, all (n = 37) of the constructed embryos developed to the Morula/blastocyst stage. When 2-cell embryos and Morulae/blastocysts were transferred into pseudopregnant females, 14 (13/96) and 24% (9/37), respectively, developed into term offspring. This study indicates that the spermatid chromosomes, which had undergone PCC, moved safely to opposite poles after oocyte activation. Since round spermatids contain no (in the mouse) or little (in patients with spermatogenic failure) oocyte-activating factor, this method may be used to rescue oocytes that fail to be activated at the time of spermatid injection.

Masato Kobayashi - One of the best experts on this subject based on the ideXlab platform.

  • oxygen consumption of single bovine embryos probed by scanning electrochemical microscopy
    Analytical Chemistry, 2001
    Co-Authors: Hitoshi Shiku, Takuo Shiraishi, Hiroaki Ohya, Tornokazu Matsue, Hiroyuki Abe, Hiroyoshi Hoshi, Masato Kobayashi
    Abstract:

    Oxygen consumption of individual bovine embryos was noninvasively quantified by scanning electrochemical microscopy (SECM). A probe microelectrode was used to scan near a single embryo surface in a culture medium to monitor the oxygen reduction current at 37 degrees C, under a water-saturated atmosphere of 5% CO2 and 95% air. The oxygen concentration profiles near the embryos were in good agreement with the theoretical spherical diffusion. When an embryo reached the stage of a Morula with a 74-microm radius on day 6 after in vitro fertilization, the oxygen concentration difference (deltaC) between the bulk solution and the Morula surface was 6.90 +/- 1.35 microM. The oxygen consumption rate (F) of the single Morula was estimated to be (1.40 +/- 0.27) x 10(-14) mol s(-1). After the SECM measurement, the embryo was continuously cultured for another 2 days and grew to the stage of a blastocyst with a 100-microm radius. For the blastocyst, the deltaC values for the inner cell mass side and the trophoblast side were 16.40 +/- 1.83 and 9.14 +/- 1.68 microM, respectively. The oxygen consumption rate of the blastocyst was found to be in the range of (2.50 +/- 0.46) x 10(-14) mol s(-1) < F < (4.49 +/- 0.50) x 10(-14) mol s(-1). We have carried out SECM measurements for 19 embryos, and the results were compared in detail with these from an optical microscopic observation. The deltaC values for the Morulae on day 6 after in vitro fertilization were strongly related to the morphological embryo quality. The Morulae showing a larger deltaC value developed into blastocysts of a larger size, and the deltaC value after the subsequent 2 days of cultivation was found to be increased.

  • oxygen consumption of single bovine embryos probed by scanning electrochemical microscopy
    Analytical Chemistry, 2001
    Co-Authors: Hitoshi Shiku, Takuo Shiraishi, Hiroaki Ohya, Tornokazu Matsue, Hiroyoshi Hoshi, Masato Kobayashi
    Abstract:

    Oxygen consumption of individual bovine embryos was noninvasively quantified by scanning electrochemical microscopy (SECM). A probe microelectrode was used to scan near a single embryo surface in a culture medium to monitor the oxygen reduction current at 37 °C, under a water-saturated atmosphere of 5% CO2 and 95% air. The oxygen concentration profiles near the embryos were in good agreement with the theoretical spherical diffusion. When an embryo reached the stage of a Morula with a 74-μm radius on day 6 after in vitro fertilization, the oxygen concentration difference (ΔC) between the bulk solution and the Morula surface was 6.90 ± 1.35 μM. The oxygen consumption rate (F) of the single Morula was estimated to be (1.40 ± 0.27) × 10-14 mol s-1. After the SECM measurement, the embryo was continuously cultured for another 2 days and grew to the stage of a blastocyst with a 100-μm radius. For the blastocyst, the ΔC values for the inner cell mass side and the trophoblast side were 16.40 ± 1.83 and 9.14 ± 1.68...

Junichiro Matsuda - One of the best experts on this subject based on the ideXlab platform.

  • birth of offspring after transfer of mongolian gerbil meriones unguiculatus embryos cryopreserved by vitrification
    Molecular Reproduction and Development, 2005
    Co-Authors: Keiji Mochida, Junichiro Matsuda, Teruhiko Wakayama, Kaoru Takano, Yoko Noguchi, Yoshie Yamamoto, Osamu Suzuki, Atsuo Ogura
    Abstract:

    The Mongolian gerbil (Meriones unguiculatus) has been used as a laboratory species in many fields of research, including neurology, oncology, and parasitology. Although the cryopreservation of embryos has become a useful means to protect valuable genetic resources, its application to the Mongolian gerbil has not yet been reported. In this study, we investigated the in vitro and in vivo developmental competence of Mongolian gerbil embryos cryopreserved by vitrification. In vivo-fertilized embryos were vitrified on the day of collection using the ethylene glycol (EG)-based solutions EFS20 and EFS40, which contained 20% and 40% EG, respectively, in PB1 containing 30% (w/v) Ficoll 70 and 0.5 M sucrose. First, we compared one-step and two-step vitrification protocols. In the one-step method, the embryos were directly transferred into the vitrification solution (EFS40), whereas in the two-step method, the embryos were exposed serially to EFS20 and EFS40 and then vitrified. After liquefying (thawing), late two-cell embryos (collected on day 3) vitrified by the two-step method showed significantly better rates of in vitro development to the Morula stage compared to those vitrified by the one-step method (65% vs. 5%, P < 0.0001). We then examined whether the same two-step method could be applied to early two-cell embryos (collected on day 2), four-cell embryos (day 4), Morulae (day 5), and blastocysts (day 6). After liquefying, 87%-100% of the embryos were morphologically normal in all groups, and 23% and 96% developed to the compacted Morula stage from early two- and four-cell embryos, respectively. After transfer into recipient females, 3% (4/123), 1% (1/102), 5% (4/73), and 10% (15/155) developed to full-term offspring from vitrified and liquefied early two-cell embryos, late two-cell embryos, Morulae, and blastocysts, respectively. This demonstrates that Mongolian gerbil embryos can be safely cryopreserved using EG-based vitrification solutions.

  • birth of offspring after transfer of mongolian gerbil meriones unguiculatus embryos cryopreserved by vitrification
    Molecular Reproduction and Development, 2005
    Co-Authors: Keiji Mochida, Junichiro Matsuda, Teruhiko Wakayama, Kaoru Takano, Yoko Noguchi, Yoshie Yamamoto, Osamu Suzuki, Atsuo Ogura
    Abstract:

    The Mongolian gerbil (Meriones unguiculatus) has been used as a laboratory species in many fields of research, including neurology, oncology, and parasitology. Although the cryopreservation of embryos has become a useful means to protect valua- ble genetic resources, its application to the Mongolian gerbil has not yet been reported. In this study, we investigated the in vitro and in vivo developmental competence of Mongolian gerbil embryos cryopre- served by vitrification. In vivo-fertilized embryos were vitrified on the day of collection using the ethylene glycol (EG)-based solutions EFS20 and EFS40, which contained 20% and 40% EG, respectively, in PB1 containing 30% (w/v) Ficoll 70 and 0.5M sucrose. First, we compared one-step and two-step vitrification protocols. In the one-step method, the embryos were directly transferred into the vitrification solution (EFS40), whereas in the two-step method, the em- bryos were exposed serially to EFS20 and EFS40 and then vitrified. After liquefying (thawing), late two-cell embryos (collected on day 3) vitrified by the two-step method showed significantly better rates of in vitro development to the Morula stage compared to those vitrified by the one-step method (65% vs. 5%, P <0.0001). We then examined whether the same two-step method could be applied to early two-cell embryos (collected on day 2), four-cell embryos (day 4), Morulae (day 5), and blastocysts (day 6). After liquefying, 87%-100% of the embryos were morpho- logically normal in all groups, and 23% and 96% developed to the compacted Morula stage from early two- and four-cell embryos, respectively. After transfer into recipient females, 3% (4/123), 1% (1/102), 5% (4/73), and 10% (15/155) developed to full-term offspring from vitrified and liquefied early two-cell embryos, late two-cell embryos, Morulae, and blas- tocysts, respectively. This demonstrates that Mongo- lian gerbil embryos can be safely cryopreserved using EG-based vitrification solutions. Mol. Reprod. Dev. 70: 464-470, 2005. 2005 Wiley-Liss, Inc.

  • mouse spermatid nuclei can support full term development after premature chromosome condensation within mature oocytes
    Human Reproduction, 1999
    Co-Authors: Atsuo Ogura, Kimiko Inoue, Junichiro Matsuda
    Abstract:

    The nucleus of round spermatids, the earliest haploid male germ cells, can participate in the formation of normal zygotes when incorporated into activated oocytes. In this study, we injected mouse round spermatids into homologous mature oocytes that were kept arrested at metaphase II to induce premature chromosome condensation (PCC) of the spermatid nuclei. After full condensation of the spermatid chromosomes, the oocytes were activated by Sr 2+ -containing medium, into which cytochalasin B was added to prevent extrusion of the segregated female and male chromosomes as polar bodies. Out of 142 oocytes examined, 104 (73%) formed two male (pseudo)pronuclei and two female pronuclei. To restore the diploid state of these zygotes, one of the female pronuclei was removed. When cultured in vitro for 72 hours, all (n = 37) of the constructed embryos developed to the Morula/blastocyst stage. When 2-cell embryos and Morulae/blastocysts were transferred into pseudopregnant females, 14 (13/96) and 24% (9/37), respectively, developed into term offspring. This study indicates that the spermatid chromosomes, which had undergone PCC, moved safely to opposite poles after oocyte activation. Since round spermatids contain no (in the mouse) or little (in patients with spermatogenic failure) oocyte-activating factor, this method may be used to rescue oocytes that fail to be activated at the time of spermatid injection.

Keiji Mochida - One of the best experts on this subject based on the ideXlab platform.

  • birth of offspring after transfer of mongolian gerbil meriones unguiculatus embryos cryopreserved by vitrification
    Molecular Reproduction and Development, 2005
    Co-Authors: Keiji Mochida, Junichiro Matsuda, Teruhiko Wakayama, Kaoru Takano, Yoko Noguchi, Yoshie Yamamoto, Osamu Suzuki, Atsuo Ogura
    Abstract:

    The Mongolian gerbil (Meriones unguiculatus) has been used as a laboratory species in many fields of research, including neurology, oncology, and parasitology. Although the cryopreservation of embryos has become a useful means to protect valua- ble genetic resources, its application to the Mongolian gerbil has not yet been reported. In this study, we investigated the in vitro and in vivo developmental competence of Mongolian gerbil embryos cryopre- served by vitrification. In vivo-fertilized embryos were vitrified on the day of collection using the ethylene glycol (EG)-based solutions EFS20 and EFS40, which contained 20% and 40% EG, respectively, in PB1 containing 30% (w/v) Ficoll 70 and 0.5M sucrose. First, we compared one-step and two-step vitrification protocols. In the one-step method, the embryos were directly transferred into the vitrification solution (EFS40), whereas in the two-step method, the em- bryos were exposed serially to EFS20 and EFS40 and then vitrified. After liquefying (thawing), late two-cell embryos (collected on day 3) vitrified by the two-step method showed significantly better rates of in vitro development to the Morula stage compared to those vitrified by the one-step method (65% vs. 5%, P <0.0001). We then examined whether the same two-step method could be applied to early two-cell embryos (collected on day 2), four-cell embryos (day 4), Morulae (day 5), and blastocysts (day 6). After liquefying, 87%-100% of the embryos were morpho- logically normal in all groups, and 23% and 96% developed to the compacted Morula stage from early two- and four-cell embryos, respectively. After transfer into recipient females, 3% (4/123), 1% (1/102), 5% (4/73), and 10% (15/155) developed to full-term offspring from vitrified and liquefied early two-cell embryos, late two-cell embryos, Morulae, and blas- tocysts, respectively. This demonstrates that Mongo- lian gerbil embryos can be safely cryopreserved using EG-based vitrification solutions. Mol. Reprod. Dev. 70: 464-470, 2005. 2005 Wiley-Liss, Inc.

  • birth of offspring after transfer of mongolian gerbil meriones unguiculatus embryos cryopreserved by vitrification
    Molecular Reproduction and Development, 2005
    Co-Authors: Keiji Mochida, Junichiro Matsuda, Teruhiko Wakayama, Kaoru Takano, Yoko Noguchi, Yoshie Yamamoto, Osamu Suzuki, Atsuo Ogura
    Abstract:

    The Mongolian gerbil (Meriones unguiculatus) has been used as a laboratory species in many fields of research, including neurology, oncology, and parasitology. Although the cryopreservation of embryos has become a useful means to protect valuable genetic resources, its application to the Mongolian gerbil has not yet been reported. In this study, we investigated the in vitro and in vivo developmental competence of Mongolian gerbil embryos cryopreserved by vitrification. In vivo-fertilized embryos were vitrified on the day of collection using the ethylene glycol (EG)-based solutions EFS20 and EFS40, which contained 20% and 40% EG, respectively, in PB1 containing 30% (w/v) Ficoll 70 and 0.5 M sucrose. First, we compared one-step and two-step vitrification protocols. In the one-step method, the embryos were directly transferred into the vitrification solution (EFS40), whereas in the two-step method, the embryos were exposed serially to EFS20 and EFS40 and then vitrified. After liquefying (thawing), late two-cell embryos (collected on day 3) vitrified by the two-step method showed significantly better rates of in vitro development to the Morula stage compared to those vitrified by the one-step method (65% vs. 5%, P < 0.0001). We then examined whether the same two-step method could be applied to early two-cell embryos (collected on day 2), four-cell embryos (day 4), Morulae (day 5), and blastocysts (day 6). After liquefying, 87%-100% of the embryos were morphologically normal in all groups, and 23% and 96% developed to the compacted Morula stage from early two- and four-cell embryos, respectively. After transfer into recipient females, 3% (4/123), 1% (1/102), 5% (4/73), and 10% (15/155) developed to full-term offspring from vitrified and liquefied early two-cell embryos, late two-cell embryos, Morulae, and blastocysts, respectively. This demonstrates that Mongolian gerbil embryos can be safely cryopreserved using EG-based vitrification solutions.

Hiroyoshi Hoshi - One of the best experts on this subject based on the ideXlab platform.

  • accumulation of cytoplasmic lipid droplets in bovine embryos and cryotolerance of embryos developed in different culture systems using serum free or serum containing media
    Molecular Reproduction and Development, 2002
    Co-Authors: Hiroyuki Abe, Shoko Yamashita, Takeshi Satoh, Hiroyoshi Hoshi
    Abstract:

    In this study, the quantitative fluctuation of cytoplasmic lipid droplets (LD) and cryotolerance were investigated in bovine embryos derived from in vitro-matured (IVM) and in vitro-fertilized (IVF) oocytes developed in different culture systems using serum-free or serum-containing media. The serum-free cultures were grown using IVMD101 medium in conjunction with bovine cumulus/granulosa cell (BCGC) cocultures or IVD101 medium without BCGC cocultures, and the serum-containing cultures were grown in the presence of BCGC cocultures using HPM199 medium supplemented with 5% calf serum (HPM199 + CS). Large numbers of sudanophilic LD were present in the cytoplasm of bovine embryos from 2-cell to hatched blastocyst stages, and the number and size differed between the embryos cultured in serum-free and serum-supplemented media. In the embryos cultured in HPM199 + CS, large (2–6 μm in diameter) sudanophilic LD increased significantly from the Morula to the blastocyst stages. Throughout the embryonic development, the embryos developed in serum-free cultures with and without BCGC cocultures had numerous sudanophilic LD, but most of these droplets were small ( 6 μm in diameter) were frequently observed in Morulae and blastocysts (including early blastocysts) developed in HPM199 + CS. Electron microscopic observations demonstrated that large LD were abundant in the cytoplasm of trophoblast and embryonic (inner cell mass) cells of blastocysts cultured in HPM199 + CS. These large LD were identified as osmophilic LD, an indication that these lipid inclusions contained a significant proportion of unsaturated lipids. Many elongated mitochondria were found in embryos developed in IVMD101 and IVD101 at the Morula and early blastocyst stages, whereas many of the mitochondria in the Morulae developed in HPM199 + CS were of an immature form such as spherical or ovoid shape. The survival and hatching rates of embryos (Morulae, early blastocysts, and blastocysts) produced in serum-free media (both IVMD101 and IVD101) after post-thaw culture were superior to those of embryos produced in serum-containing medium. These results showed that bovine embryos cultured in serum-containing medium abnormally accumulated cytoplasmic lipids into their cytoplasm and the excess accumulation of cytoplasmic LD in embryos may affect the cryotolerance of embryos. Mol. Reprod. Dev. 61: 57–66, 2002. © 2002 Wiley-Liss, Inc.

  • oxygen consumption of single bovine embryos probed by scanning electrochemical microscopy
    Analytical Chemistry, 2001
    Co-Authors: Hitoshi Shiku, Takuo Shiraishi, Hiroaki Ohya, Tornokazu Matsue, Hiroyuki Abe, Hiroyoshi Hoshi, Masato Kobayashi
    Abstract:

    Oxygen consumption of individual bovine embryos was noninvasively quantified by scanning electrochemical microscopy (SECM). A probe microelectrode was used to scan near a single embryo surface in a culture medium to monitor the oxygen reduction current at 37 degrees C, under a water-saturated atmosphere of 5% CO2 and 95% air. The oxygen concentration profiles near the embryos were in good agreement with the theoretical spherical diffusion. When an embryo reached the stage of a Morula with a 74-microm radius on day 6 after in vitro fertilization, the oxygen concentration difference (deltaC) between the bulk solution and the Morula surface was 6.90 +/- 1.35 microM. The oxygen consumption rate (F) of the single Morula was estimated to be (1.40 +/- 0.27) x 10(-14) mol s(-1). After the SECM measurement, the embryo was continuously cultured for another 2 days and grew to the stage of a blastocyst with a 100-microm radius. For the blastocyst, the deltaC values for the inner cell mass side and the trophoblast side were 16.40 +/- 1.83 and 9.14 +/- 1.68 microM, respectively. The oxygen consumption rate of the blastocyst was found to be in the range of (2.50 +/- 0.46) x 10(-14) mol s(-1) < F < (4.49 +/- 0.50) x 10(-14) mol s(-1). We have carried out SECM measurements for 19 embryos, and the results were compared in detail with these from an optical microscopic observation. The deltaC values for the Morulae on day 6 after in vitro fertilization were strongly related to the morphological embryo quality. The Morulae showing a larger deltaC value developed into blastocysts of a larger size, and the deltaC value after the subsequent 2 days of cultivation was found to be increased.

  • oxygen consumption of single bovine embryos probed by scanning electrochemical microscopy
    Analytical Chemistry, 2001
    Co-Authors: Hitoshi Shiku, Takuo Shiraishi, Hiroaki Ohya, Tornokazu Matsue, Hiroyoshi Hoshi, Masato Kobayashi
    Abstract:

    Oxygen consumption of individual bovine embryos was noninvasively quantified by scanning electrochemical microscopy (SECM). A probe microelectrode was used to scan near a single embryo surface in a culture medium to monitor the oxygen reduction current at 37 °C, under a water-saturated atmosphere of 5% CO2 and 95% air. The oxygen concentration profiles near the embryos were in good agreement with the theoretical spherical diffusion. When an embryo reached the stage of a Morula with a 74-μm radius on day 6 after in vitro fertilization, the oxygen concentration difference (ΔC) between the bulk solution and the Morula surface was 6.90 ± 1.35 μM. The oxygen consumption rate (F) of the single Morula was estimated to be (1.40 ± 0.27) × 10-14 mol s-1. After the SECM measurement, the embryo was continuously cultured for another 2 days and grew to the stage of a blastocyst with a 100-μm radius. For the blastocyst, the ΔC values for the inner cell mass side and the trophoblast side were 16.40 ± 1.83 and 9.14 ± 1.68...