title: Long-term live-cell imaging of mammalian preimplantation development and derivation process of pluripotent stem cells from the embryos.
abstract: Mammalian fertilization is a process in which two highly specialized haploid gametes unite and endow totipotency to the resulting diploid zygote. This is followed by cell proliferation and the onset of differentiation during the brief period leading up to implantation. In these processes, a number of cellular components and structures are regulated spatially and temporally, as seen in repeated cell division, cell cycle progression, and epigenetic reprogramming. In mammals, the numbers of oocytes and embryos that can be collected are very limited. Therefore, analyses of molecular mechanisms are hampered because of difficulties in conducting biochemical analyses on such limited material. Furthermore, immunostaining methods require cell fixation and are insufficient for understanding ontogeny, because the processes observed in fertilization and early embryonic development progress in time-dependent manners and each phenomenon is connected with others by cause-and-effect relationships. Consequently, it is important to develop an experimental system that enables molecular imaging without affecting embryonic development. To achieve the above advantages, especially retrospective and prospective analyses, we have established a live-cell imaging system that enables observations under minimally invasive conditions. Using this approach, we have succeeded in visualizing and predicting the developmental potential of embryos after various perturbations. We also succeeded in imaging embryonic stem (ES) cell derivation in natural conditions. In this review, we describe a brief history of embryonic imaging and detailed protocols. We also discuss promising aspects of imaging in the fields of developmental and stem cell biology.
authors.affiliation: Center for Genetic Analysis of Biological Responses, Research Institute for Microbial Diseases, Osaka University, Suita, Japan. yamagata@biken.osaka-u.ac.jp
authors.forename: Kazuo
authors.initials: K
authors.lastname: Yamagata
authors.forename: Jun
authors.initials: J
authors.lastname: Ueda
mesh_terms: D000818:Animals; D001755:Blastocyst / Q000166:cytology*; D002453:Cell Cycle; D002455:Cell Division; D053595:Embryonic Stem Cells / Q000166:cytology; D004867:Equipment Design; D005260:Female; D005307:Fertilization in Vitro; D049452:Green Fluorescent Proteins / Q000378:metabolism; D051379:Mice; D018613:Microscopy, Confocal; D009865:Oocytes / Q000166:cytology; D039904:Pluripotent Stem Cells / Q000166:cytology*; D012313:RNA / Q000378:metabolism
chemical_list: D049452:Green Fluorescent Proteins; D012313:RNA