Abstract: A new prenatal genetic testing technique using cell-free fetal DNA in maternal plasma, the Non-Invasive Prenatal Test (or NIPT), was introduced in Japan in 2013. The NIPT is easy and safe but not definitive; it also has high sensitivity and specificity under certain conditions. In the near future, it could be used not only for detecting chromosomal aneuploidy, but also complete genome analysis of the fetus. However, termination of pregnancy is the only option for those wanting to avoid having a baby with a chromosomal anomaly or a genetic disease. Given this situation, this review explores the ethical and social issues surrounding prenatal genetic testing focusing on three points: 1) selective abortion as an ethical issue, 2) informed consent and decision making, and 3) alternative perspectives on prenatal testing.
{"title":"Ethical and Social Implications of Current Prenatal Genetic Testing","authors":"Azumi Tsuge","doi":"10.1274/jmor.33.109","DOIUrl":"https://doi.org/10.1274/jmor.33.109","url":null,"abstract":"Abstract: A new prenatal genetic testing technique using cell-free fetal DNA in maternal plasma, the Non-Invasive Prenatal Test (or NIPT), was introduced in Japan in 2013. The NIPT is easy and safe but not definitive; it also has high sensitivity and specificity under certain conditions. In the near future, it could be used not only for detecting chromosomal aneuploidy, but also complete genome analysis of the fetus. However, termination of pregnancy is the only option for those wanting to avoid having a baby with a chromosomal anomaly or a genetic disease. Given this situation, this review explores the ethical and social issues surrounding prenatal genetic testing focusing on three points: 1) selective abortion as an ethical issue, 2) informed consent and decision making, and 3) alternative perspectives on prenatal testing.","PeriodicalId":90599,"journal":{"name":"Journal of mammalian ova research","volume":"73 1","pages":"109 - 113"},"PeriodicalIF":0.0,"publicationDate":"2016-10-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"79602907","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Abstract: Mitochondrial DNA (mtDNA) mutation is associated with serious human disorders and affects multiple organs and tissues with high-energy requirements. Since the transmission of mtDNA is complex and is not fully understood, an accurate estimation of mtDNA disease transmission by preimplantation genetic diagnosis (PGD) or by prenatal diagnosis (PND) remains challenging. Recently, nuclear transfer techniques, including maternal spindle transfer (MST), pronuclear transfer (PNT) and polar body transfer (PBT), have shown the promising results. These methods avoid the transmission of mutated mtDNA from mother to offspring, and are collectively known as the mitochondrial replacement therapy (MRT). Further, the United Kingdom Parliament approved the Human Fertilisation and Embryology Authority (HFEA) to grant licenses for experimental use of MST and PNT in humans in 2015. Thus, a new era of assisted reproductive technology (ART), in which cures can be provided at the gamete or early zygote stages, is realistically approaching. In this review, we summarize the methods and the challenges confronting the clinical application of MRT.
{"title":"Challenges Towards Establishing Germline Gene Therapy for Inherited Mitochondrial Diseases","authors":"Naomi Shiga, Masahito Tachibana, N. Yaegashi","doi":"10.1274/jmor.33.89","DOIUrl":"https://doi.org/10.1274/jmor.33.89","url":null,"abstract":"Abstract: Mitochondrial DNA (mtDNA) mutation is associated with serious human disorders and affects multiple organs and tissues with high-energy requirements. Since the transmission of mtDNA is complex and is not fully understood, an accurate estimation of mtDNA disease transmission by preimplantation genetic diagnosis (PGD) or by prenatal diagnosis (PND) remains challenging. Recently, nuclear transfer techniques, including maternal spindle transfer (MST), pronuclear transfer (PNT) and polar body transfer (PBT), have shown the promising results. These methods avoid the transmission of mutated mtDNA from mother to offspring, and are collectively known as the mitochondrial replacement therapy (MRT). Further, the United Kingdom Parliament approved the Human Fertilisation and Embryology Authority (HFEA) to grant licenses for experimental use of MST and PNT in humans in 2015. Thus, a new era of assisted reproductive technology (ART), in which cures can be provided at the gamete or early zygote stages, is realistically approaching. In this review, we summarize the methods and the challenges confronting the clinical application of MRT.","PeriodicalId":90599,"journal":{"name":"Journal of mammalian ova research","volume":"5 1","pages":"89 - 99"},"PeriodicalIF":0.0,"publicationDate":"2016-10-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"88482883","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
K. Kyono, T. Hashimoto, M. Kuchiki, E. Sakamoto, Masayo Tani, Atsuko Tanaka, Sachiko Nonaka, Yusuke Nakamura, Y. Nakajo, N. Aono, T. Takeuchi
Abstract: Purpose: To highlight the imperative of informed consent in the fertility preservation of cancer patients before ovarian tissue autotransplantation. Methods: Three papers of tumor recurrence after autotransplantation of frozen-thawed ovarian tissue were compared with the main papers before tumor recurrence was reported in the cancer patients. Results: Histology was performed before autotransplantation in cases 1 and 3, but not in case 2. Histology alone is insufficient for the detection of minimal residual disease (MRD). Furthermore, transplanted ovarian tissue is different from that examined for MRD detection, and how much of the resected ovarian tissue was examined for MRD detection is unclear. The possibility that grafted tissue caused tumor recurrence cannot be ruled out in any of the three cases, because autotransplantation in cancer patients, at present, uses ovarian tissue different from that examined for MRD. Conclusions: We recommend informed consent for cancer patients, because: 1) the transplanted ovarian tissue is different from the ovarian tissue examined for MRD detection; 2) the amount of resected ovarian tissue analyzed for MRD is very small; and 3) MRD detection methods vary. In conclusion, freezing and storage of ovaries should be encouraged, transplantation must be performed carefully, and informed consent is essential.
{"title":"A Review: the Essentials of Informed Consent for Cancer Patients Before Autologous Transplantation of Ovarian Tissue","authors":"K. Kyono, T. Hashimoto, M. Kuchiki, E. Sakamoto, Masayo Tani, Atsuko Tanaka, Sachiko Nonaka, Yusuke Nakamura, Y. Nakajo, N. Aono, T. Takeuchi","doi":"10.1274/jmor.33.115","DOIUrl":"https://doi.org/10.1274/jmor.33.115","url":null,"abstract":"Abstract: Purpose: To highlight the imperative of informed consent in the fertility preservation of cancer patients before ovarian tissue autotransplantation. Methods: Three papers of tumor recurrence after autotransplantation of frozen-thawed ovarian tissue were compared with the main papers before tumor recurrence was reported in the cancer patients. Results: Histology was performed before autotransplantation in cases 1 and 3, but not in case 2. Histology alone is insufficient for the detection of minimal residual disease (MRD). Furthermore, transplanted ovarian tissue is different from that examined for MRD detection, and how much of the resected ovarian tissue was examined for MRD detection is unclear. The possibility that grafted tissue caused tumor recurrence cannot be ruled out in any of the three cases, because autotransplantation in cancer patients, at present, uses ovarian tissue different from that examined for MRD. Conclusions: We recommend informed consent for cancer patients, because: 1) the transplanted ovarian tissue is different from the ovarian tissue examined for MRD detection; 2) the amount of resected ovarian tissue analyzed for MRD is very small; and 3) MRD detection methods vary. In conclusion, freezing and storage of ovaries should be encouraged, transplantation must be performed carefully, and informed consent is essential.","PeriodicalId":90599,"journal":{"name":"Journal of mammalian ova research","volume":"29 1","pages":"115 - 118"},"PeriodicalIF":0.0,"publicationDate":"2016-10-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"88551800","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Abstract: The survival probability of mouse blastocysts developed from nuclear transfer (NT) was estimated prior to embryo transfer using a novel biopsy method. NT embryos were separated into two at the 2-cell stage to produce monozygotic twin blastocysts from a single NT oocyte. We first examined efficient culture methods to produce monozygotic twin blastocysts. Then one of the two blastocysts was used for gene expression analysis, and the other was transferred to a recipient mouse. Co-culture of monozygotic twin blastomeres with fertilized embryos or culture in a small well in vitro improved the developmental potential to the blastocyst stage and increased the blastocyst cell number. Although 21.9% of twin somatic cell NT embryos had the same expression levels of Oct4 and Sox2 genes as fertilized embryos, no fetuses were obtained after transfer to recipients.
{"title":"Estimating the Survival Probability of Nuclear-Transfer Embryos before Embryo Transfer by a Novel Biopsy: Oct4 and Sox2 Gene Expression Patterns of a Monozygotic Twin Blastocyst Separated at the 2-Cell Stage of Nuclear-Transfer Embryos","authors":"Kazuki Ohata, Y. Kato","doi":"10.1274/jmor.33.55","DOIUrl":"https://doi.org/10.1274/jmor.33.55","url":null,"abstract":"Abstract: \u0000 The survival probability of mouse blastocysts developed from nuclear transfer (NT) was estimated prior to embryo transfer using a novel biopsy method. NT embryos were separated into two at the 2-cell stage to produce monozygotic twin blastocysts from a single NT oocyte. We first examined efficient culture methods to produce monozygotic twin blastocysts. Then one of the two blastocysts was used for gene expression analysis, and the other was transferred to a recipient mouse. Co-culture of monozygotic twin blastomeres with fertilized embryos or culture in a small well in vitro improved the developmental potential to the blastocyst stage and increased the blastocyst cell number. Although 21.9% of twin somatic cell NT embryos had the same expression levels of Oct4 and Sox2 genes as fertilized embryos, no fetuses were obtained after transfer to recipients.","PeriodicalId":90599,"journal":{"name":"Journal of mammalian ova research","volume":"11 1","pages":"55 - 61"},"PeriodicalIF":0.0,"publicationDate":"2016-05-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"85792652","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Abstract: The TYH medium was first reported as a medium for in vitro fertilization (IVF) of mouse eggs with epididymal spermatozoa, by Toyoda, Yokoyama and Hosi, in 1971. It was a modified Krebs-Ringer bicarbonate solution, supplemented with glucose, Na-pyruvate, antibiotics and bovine serum albumin. In TYH medium, almost all eggs are fertilized within 1 h, if the spermatozoa are pre-incubated for 2 h before insemination, while the sperm penetration is delayed for approximately 1 h when fresh epididymal spermatozoa are used. These findings showed that sperm capacitation can be induced in a chemically defined medium without requiring the female reproductive tract. Although the medium was not specifically named in the original paper, it was later called “TYH” after the initials of the three authors of the original paper. The IVF method using TYH medium is widely used due to its high reproducibility. In this mini-review, we describe the early efforts to develop the TYH medium and briefly discuss the related areas.
{"title":"The Early History of the TYH Medium for in vitro Fertilization of Mouse Ova","authors":"Y. Toyoda, M. Yokoyama","doi":"10.1274/jmor.33.3","DOIUrl":"https://doi.org/10.1274/jmor.33.3","url":null,"abstract":"Abstract: The TYH medium was first reported as a medium for in vitro fertilization (IVF) of mouse eggs with epididymal spermatozoa, by Toyoda, Yokoyama and Hosi, in 1971. It was a modified Krebs-Ringer bicarbonate solution, supplemented with glucose, Na-pyruvate, antibiotics and bovine serum albumin. In TYH medium, almost all eggs are fertilized within 1 h, if the spermatozoa are pre-incubated for 2 h before insemination, while the sperm penetration is delayed for approximately 1 h when fresh epididymal spermatozoa are used. These findings showed that sperm capacitation can be induced in a chemically defined medium without requiring the female reproductive tract. Although the medium was not specifically named in the original paper, it was later called “TYH” after the initials of the three authors of the original paper. The IVF method using TYH medium is widely used due to its high reproducibility. In this mini-review, we describe the early efforts to develop the TYH medium and briefly discuss the related areas.","PeriodicalId":90599,"journal":{"name":"Journal of mammalian ova research","volume":"1 1","pages":"10 - 3"},"PeriodicalIF":0.0,"publicationDate":"2016-05-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"82044649","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Abstract: Systematic studies of mouse embryo culture beginning in 1949 led to an understanding of essential medium components for early mammalian embryos, and embryo culture from the zygote to the blastocyst stage was achieved in 1968. Since then, medium components that are either beneficial or detrimental for embryo culture have been identified. A variety of culture media that mimic the female reproductive tract, such as human tubal fluid medium and sequential media, were developed from the 1970s to the 1990s, and a single medium in which the concentrations of components were determined by a simplex optimization method was introduced for clinical use in 2002. While either sequential media or a single medium is currently used in most cases, no conclusion has yet been reached as to which of the two approaches is the best. That we are now easily able to culture embryos is the result of the work of pioneers. This review presents a chronological overview of media development from initial attempts at mouse embryo culture using synthetic media to the human embryo culture media used today. It also presents the characteristics of sequential media and a single medium. Finally, problems observed with current embryo culture media are discussed, along with future development in this area.
{"title":"Human Preimplantation Embryo Culture Media: Past, Present, and Future","authors":"Tatsuma Yao, Yuta Asayama","doi":"10.1274/jmor.33.17","DOIUrl":"https://doi.org/10.1274/jmor.33.17","url":null,"abstract":"Abstract: \u0000 Systematic studies of mouse embryo culture beginning in 1949 led to an understanding of essential medium components for early mammalian embryos, and embryo culture from the zygote to the blastocyst stage was achieved in 1968. Since then, medium components that are either beneficial or detrimental for embryo culture have been identified. A variety of culture media that mimic the female reproductive tract, such as human tubal fluid medium and sequential media, were developed from the 1970s to the 1990s, and a single medium in which the concentrations of components were determined by a simplex optimization method was introduced for clinical use in 2002. While either sequential media or a single medium is currently used in most cases, no conclusion has yet been reached as to which of the two approaches is the best. That we are now easily able to culture embryos is the result of the work of pioneers. This review presents a chronological overview of media development from initial attempts at mouse embryo culture using synthetic media to the human embryo culture media used today. It also presents the characteristics of sequential media and a single medium. Finally, problems observed with current embryo culture media are discussed, along with future development in this area.","PeriodicalId":90599,"journal":{"name":"Journal of mammalian ova research","volume":"189 1","pages":"17 - 34"},"PeriodicalIF":0.0,"publicationDate":"2016-05-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"86233093","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Abstract: Although vitrification and follicular survival subsequent to xenotransplantation of canine ovarian tissues have been reported, the degree of cryoinjury after the vitrification has not yet been fully examined. Since cryoinjury after vitrification of ovaries has been reported in bovine and humans, this study evaluated canine ovarian tissues after vitrification by immunohistochemistry and TUNEL assay. Ovaries were collected from immature bitches. The ovarian tissue cubes were pretreated with 1 M DMSO. Subsequently, DAP 213 solution (2 M DMSO, 1 M acetamide, 3 M propylene glycol) was added. The cryotubes containing the ovarian tissues were placed on ice for 5 min, then plunged directly into liquid nitrogen. After warming with 0.25 M sucrose, cryopreserved ovarian tissues were fixed and subsequently stained with hematoxylin-eosin, proliferation cell nuclear antigen (PCNA) antibody, or active caspase-3 (AC-3) antibody, and evaluated by the TUNEL assay. There were no differences in the numbers of primordial, primary, secondary and antral follicles/mm2 between the fresh and vitrified-warmed ovarian tissues. Percentages of PCNA and AC-3 antibody positive cells were similar regardless of the developmental stage of the follicles and experimental group. A few TUNEL positive cells were detected in both experimental groups. These results suggest that vitrification with DAP213 does not induce cryoinjury in ovarian tissues from immature bitches.
摘要:尽管已经有报道报道了犬卵巢组织异种移植后的玻璃化和卵泡存活，但玻璃化后的冷冻损伤程度尚未得到充分的研究。由于在牛和人的卵巢玻璃化后冷冻损伤的报道，本研究通过免疫组织化学和TUNEL试验评估了玻璃化后的犬卵巢组织。从未成熟母狗身上采集卵巢。用1 M DMSO预处理卵巢组织块。随后加入DAP 213溶液(2 M DMSO, 1 M乙酰胺，3 M丙二醇)。将含有卵巢组织的冷冻管置于冰上5分钟，然后直接放入液氮中。用0.25 M蔗糖加热后，冷冻保存的卵巢组织固定，随后用苏木精-伊红、增殖细胞核抗原(PCNA)抗体或活性caspase-3 (AC-3)抗体染色，并通过TUNEL试验进行评价。新鲜卵巢组织和玻璃化卵巢组织的原始卵泡、初级卵泡、次级卵泡和窦卵泡的数量/mm2没有差异。无论卵泡和实验组的发育阶段如何，PCNA和AC-3抗体阳性细胞的百分比相似。两组均有少量TUNEL阳性细胞。这些结果表明，用DAP213玻璃化处理不会引起未成熟母狗卵巢组织的冷冻损伤。
{"title":"Incidence of Apoptotic Cells After Vitrification in Canine Ovarian Tissues","authors":"Madoka Hariya, Hiroshi Suzuki","doi":"10.1274/jmor.33.69","DOIUrl":"https://doi.org/10.1274/jmor.33.69","url":null,"abstract":"Abstract: Although vitrification and follicular survival subsequent to xenotransplantation of canine ovarian tissues have been reported, the degree of cryoinjury after the vitrification has not yet been fully examined. Since cryoinjury after vitrification of ovaries has been reported in bovine and humans, this study evaluated canine ovarian tissues after vitrification by immunohistochemistry and TUNEL assay. Ovaries were collected from immature bitches. The ovarian tissue cubes were pretreated with 1 M DMSO. Subsequently, DAP 213 solution (2 M DMSO, 1 M acetamide, 3 M propylene glycol) was added. The cryotubes containing the ovarian tissues were placed on ice for 5 min, then plunged directly into liquid nitrogen. After warming with 0.25 M sucrose, cryopreserved ovarian tissues were fixed and subsequently stained with hematoxylin-eosin, proliferation cell nuclear antigen (PCNA) antibody, or active caspase-3 (AC-3) antibody, and evaluated by the TUNEL assay. There were no differences in the numbers of primordial, primary, secondary and antral follicles/mm2 between the fresh and vitrified-warmed ovarian tissues. Percentages of PCNA and AC-3 antibody positive cells were similar regardless of the developmental stage of the follicles and experimental group. A few TUNEL positive cells were detected in both experimental groups. These results suggest that vitrification with DAP213 does not induce cryoinjury in ovarian tissues from immature bitches.","PeriodicalId":90599,"journal":{"name":"Journal of mammalian ova research","volume":"24 1","pages":"69 - 75"},"PeriodicalIF":0.0,"publicationDate":"2016-05-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"84561695","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Abstract: Rat 1-cell embryos fertilized in vivo or in vitro show a complete developmental block at the 2- to 4-cell stage in conventional culture media. To develop a culture medium supporting early embryonic development in rats, we examined the effects of some chemical and physical factors on the development of rat 1-cell embryos. The results indicated that (1) the developmental block is caused by phosphate, (2) glucose and amino acids are necessary, (3) the optimal osmolarity is 244–246 mOsm, and (4) the presence of bovine serum albumin and relatively high (110.0–130.0 mM) NaCl concentrations in the early stages preceding pronucleus formation is necessary. On the basis of these results, novel culture media designated as modified rat 1-cell embryo culture medium (mR1ECM) and mR1ECM for in vitro fertilization (mR1ECM-IVF) were developed. In cultures using these media, 90% or more of rat 1-cell embryos fertilized in vivo or in vitro successfully developed to the blastocyst stage in vitro.
{"title":"Development of a Culture Medium for Rat 1-Cell Embryos","authors":"K. Miyoshi","doi":"10.1274/jmor.33.11","DOIUrl":"https://doi.org/10.1274/jmor.33.11","url":null,"abstract":"Abstract: Rat 1-cell embryos fertilized in vivo or in vitro show a complete developmental block at the 2- to 4-cell stage in conventional culture media. To develop a culture medium supporting early embryonic development in rats, we examined the effects of some chemical and physical factors on the development of rat 1-cell embryos. The results indicated that (1) the developmental block is caused by phosphate, (2) glucose and amino acids are necessary, (3) the optimal osmolarity is 244–246 mOsm, and (4) the presence of bovine serum albumin and relatively high (110.0–130.0 mM) NaCl concentrations in the early stages preceding pronucleus formation is necessary. On the basis of these results, novel culture media designated as modified rat 1-cell embryo culture medium (mR1ECM) and mR1ECM for in vitro fertilization (mR1ECM-IVF) were developed. In cultures using these media, 90% or more of rat 1-cell embryos fertilized in vivo or in vitro successfully developed to the blastocyst stage in vitro.","PeriodicalId":90599,"journal":{"name":"Journal of mammalian ova research","volume":"54 10","pages":"11 - 16"},"PeriodicalIF":0.0,"publicationDate":"2016-05-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1274/jmor.33.11","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"72507847","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Mitsutoshi Yamada, T. Hamatani, H. Akutsu, Mamoru Tanaka
Abstract: Assisted reproductive technology (ART) has greatly benefited numerous infertile couples who would never have had their babies without this technique. However, in vitro culture is reported to cause epigenetic and transcriptomic changes on preimplantation embryos, leading to adverse effect on development, and little is known about the molecular mechanisms underlying these changes. Here, we first introduce key studies that designate the effectiveness of an omic strategy to explore the molecular mechanisms governing preimplantation development of in vitro-cultured embryos. Furthermore, we review how in vitro culture components facilitate genomic reprogramming and zygotic genome activation (ZGA) contributing to preimplantation development after somatic cell nuclear transfer (SCNT). From these different perspectives, we would search for a breakthrough to further optimize preimplantation embryo culture conditions and improve ART.
{"title":"Towards Further Optimization of Preimplantation Embryo Culture Media: from the Viewpoint of Omics and Somatic Cell Nuclear Transfer (SCNT) Studies","authors":"Mitsutoshi Yamada, T. Hamatani, H. Akutsu, Mamoru Tanaka","doi":"10.1274/jmor.33.35","DOIUrl":"https://doi.org/10.1274/jmor.33.35","url":null,"abstract":"Abstract: \u0000 Assisted reproductive technology (ART) has greatly benefited numerous infertile couples who would never have had their babies without this technique. However, in vitro culture is reported to cause epigenetic and transcriptomic changes on preimplantation embryos, leading to adverse effect on development, and little is known about the molecular mechanisms underlying these changes. Here, we first introduce key studies that designate the effectiveness of an omic strategy to explore the molecular mechanisms governing preimplantation development of in vitro-cultured embryos. Furthermore, we review how in vitro culture components facilitate genomic reprogramming and zygotic genome activation (ZGA) contributing to preimplantation development after somatic cell nuclear transfer (SCNT). From these different perspectives, we would search for a breakthrough to further optimize preimplantation embryo culture conditions and improve ART.","PeriodicalId":90599,"journal":{"name":"Journal of mammalian ova research","volume":"17 1","pages":"35 - 43"},"PeriodicalIF":0.0,"publicationDate":"2016-05-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"85340478","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Abstract: Mouse parthenogenetic (PG) embryos do not survive beyond day 9.5 of pregnancy. In this study, to understand the molecular mechanisms underlying the failure of parthenogenesis at the early developmental stage, we performed global gene expression profiling of the PG inner cell mass (ICM) and trophectoderm (TE) using microarray analysis, compared the results with those from in vitro-fertilized embryos, and identified genes whose expression levels showed more than a 4-fold change as a cutoff. Eighty probe sets were up-regulated and 59 probe sets down-regulated in the PG ICM, while 169 and 43 probe sets were respectively up-regulated and downregulated in PG TE. We selected two genes (Sfmbt2 and Gab1) that were down-regulated in both the PG ICM and TE, one gene (Nat1) that was down-regulated in the PG ICM, and one gene (Lysmd2) that was up-regulated in the PG TE, and analyzed the gene expression levels using real-time PCR. The quantitative expression levels of these four genes were confirmed by real-time PCR. In the present study, we identified differentially expressed genes in PG embryos and also identified those that were ICM- or TE-specific in PG embryos.
{"title":"Microarray Analysis of Differentially Expressed Genes in Inner Cell Mass and Trophectoderm of Parthenogenetic Embryos","authors":"Shiori Goto, F. Cao, T. Kono, H. Ogawa","doi":"10.1274/jmor.33.45","DOIUrl":"https://doi.org/10.1274/jmor.33.45","url":null,"abstract":"Abstract: \u0000 Mouse parthenogenetic (PG) embryos do not survive beyond day 9.5 of pregnancy. In this study, to understand the molecular mechanisms underlying the failure of parthenogenesis at the early developmental stage, we performed global gene expression profiling of the PG inner cell mass (ICM) and trophectoderm (TE) using microarray analysis, compared the results with those from in vitro-fertilized embryos, and identified genes whose expression levels showed more than a 4-fold change as a cutoff. Eighty probe sets were up-regulated and 59 probe sets down-regulated in the PG ICM, while 169 and 43 probe sets were respectively up-regulated and downregulated in PG TE. We selected two genes (Sfmbt2 and Gab1) that were down-regulated in both the PG ICM and TE, one gene (Nat1) that was down-regulated in the PG ICM, and one gene (Lysmd2) that was up-regulated in the PG TE, and analyzed the gene expression levels using real-time PCR. The quantitative expression levels of these four genes were confirmed by real-time PCR. In the present study, we identified differentially expressed genes in PG embryos and also identified those that were ICM- or TE-specific in PG embryos.","PeriodicalId":90599,"journal":{"name":"Journal of mammalian ova research","volume":"16 1","pages":"45 - 54"},"PeriodicalIF":0.0,"publicationDate":"2016-05-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"87634244","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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