The Culture of Preimplantation Rat Embryos and the Production of Allophenic Rats

Mayer, Jacob; Fritz, Hans

doi:10.1530/jrf.0.0390001

Cited by 62 publications

(31 citation statements)

References 7 publications

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“…Gerbil preimplantation embryos may require pyruvate and lactate in the culture systems, since the development of rat 1-cell embryos to blastocysts was enhanced by the addition of pyruvate and lactate to the medium [4]. Developmental blocks in vitro were reported in rat [25] and hamster [26,27] embryos at the 2 to 4-cell stages, in outbred strains of mice at the 2-cell stage [28,29], in sheep [30] and cow [30][31][32] embryos at the 8-16-cell stage, and in pig embryos at the 4-cell stage [33][34][35]. In the present study, 1-cell gerbil embryos were blocked at the 16-cell stage when they were cultured in mTCM199 without oviductal cells.…”

Section: Discussionmentioning

confidence: 99%

Development of Mongolian Gerbil 1-cell Embryos to Blastocysts in Co-culture with Oviductal Cells in TCM 199 Supplemented with Pyruvate, Lactate and Fetal Calf Serum.

Tsujii

Saruwatari

Hamano

2002

Journal of Reproduction and Development

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Abstract.To develop an in vitro culture system for Mongolian gerbil embryos, we examined the effect of co-culture with oviductal epithelial cells on the development of 1-and 2-cell embryos collected from superovulated gerbils. In the first experiment, 2-cell embryos were cultured in modified TCM 199 supplemented with pyruvate, lactate and fetal calf serum (mTCM199) with or without gerbil or mouse oviductal cells. In the second experiment, in 1-cell embryos were cultured with or without gerbil oviductal cells in mTCM199. Although, 2-cell gerbil embryos did not develop beyond the 16-cell stage without oviductal cells in both TCM199 and mTCM199, co-culture with gerbil or mouse oviductal cells supported the development of 2-cell gerbil embryos to blastocysts. Co-culture with gerbil oviductal cells in mTCM199, but not in TCM199, also supported the development of 1-cell embryos to blastocysts. There were no differences between the in vivo and in vitro developed blastocysts in morphological appearance and cell numbers. This co-culture system can be used for in vitro culture of Mongolian gerbil embryos.

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Section: Discussionmentioning

confidence: 99%

Development of Mongolian Gerbil 1-cell Embryos to Blastocysts in Co-culture with Oviductal Cells in TCM 199 Supplemented with Pyruvate, Lactate and Fetal Calf Serum.

Tsujii

Saruwatari

Hamano

2002

Journal of Reproduction and Development

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“…The process of cell lineage specification in the preimplantation embryo, and how and when developmental potential becomes restricted, have been the subject of intense scrutiny over the last ten years. The existence of chimaeras generated by the combination of cells from different embryos provides strong evidence for the regulative ability of the early mammalian embryo (Gardner, 1968; Gardner and Munro, 1974;Mayer and Fritz, 1974;Tucker et al, 1974; Fehilly et al, 1984b; Fehilly et al, 1984a;Tachibana et al, 2012). Chimaeras occur naturally during the development of marmosets (Gengozian et al, 1964;Ross et al, 2007) and their existence in humans has been dramatically revealed in various court cases and paternity suits (Ainsworth, 2003).…”

Section: Assessing Developmental Potential: Merging and Splitting Embmentioning

confidence: 99%

A molecular basis for developmental plasticity in early mammalian embryos

2013

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SummaryEarly mammalian embryos exhibit remarkable plasticity, as highlighted by the ability of separated early blastomeres to produce a whole organism. Recent work in the mouse implicates a network of transcription factors in governing the establishment of the primary embryonic lineages. A combination of genetics and embryology has uncovered the organisation and function of the components of this network, revealing a gradual resolution from ubiquitous to lineage-specific expression through a combination of defined regulatory relationships, spatially organised signalling, and biases from mechanical inputs. Here, we summarise this information, link it to classical embryology and propose a molecular framework for the establishment and regulation of developmental plasticity. Key words: Chimaera, Developmental plasticity, Regulative development, Stochastic gene expression, Twin IntroductionOne of the most intriguing observations in developmental biology was reported by Hans Driesch in 1892 when testing the dogma of the time, which had been established by W. Roux (Roux, 1888; see Sander, 1991), that potencies, or 'prospective cell fates' (see Glossary, Box 1) as we call them today, are progressively and irreversibly restricted from the first cleavage of an embryo (Driesch, 1892). Driesch established a clean experimental protocol to split the early blastomeres of sea urchin embryos and analyse their fates during development. Not without surprise he observed that, upon separation, individual blastomeres from the 2-and 4-cell stages could give rise to a complete sea urchin larva (Fig. 1A). This indicated that the fates of the first blastomeres were not fixed, as had been suggested by Roux, but exhibited a large degree of plasticity (see Glossary, Box 1), i.e. the blastomeres were totipotent (Sander, 1991;Sander, 1992). As a result of these experiments he could experimentally induce twins and quadruplets. Similar results were obtained through related experiments in other embryos, including frogs (Morgan, 1895) -which had been the subject of Roux's work -revealing that the full developmental potential of the zygote (totipotency, see Glossary, Box 1) is maintained through at least the first divisions of the embryo.These experiments highlight a transient maintenance of developmental potential (see Glossary, Box 1), which is not restricted to the early stages of development, and indicate that, within an embryo, the potential of a cell or group of cells is greater than its actual fate (Fig. 1B) (Wolpert and Tickle, 2011). Furthermore, this potential can be captured and replicated, as in the Development 140, 3499-3510 (2013) HYPOTHESIS Box 1. GlossaryCell fate. The developmental destination of a cell if left undisturbed in its environment. It is revealed through lineagetracing experiments in which a cell is labelled and its progeny followed. The fate of a cell is more restricted than its potential. Cell state. A transient condition with a variable degree of stability; a stepping stone in the chain that configures a path to ...

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“…The embryos were equilibrated with 1-5 M-dimethylsulphoxide (DMSO) at 0°C for 15 min by the addition of a further 0-1 ml PB1 containing 3 m-DMSO. The procedures for freezing and thawing were similar to those used for mouse embryos and have been described elsewhere (Whittingham et Whittingham, 1974b (Mayer & Fritz, 1974;Whittingham, 1975). In the mouse, there is a very close approximation between the number of embryos morphologically normal at recovery and the number that continue development to the blastocyst stage in vitro (Whittingham et al, 1972).…”

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confidence: 99%

Survival of Rat Embryos After Freezing and Thawing

Whittingham¹

1975

Reproduction

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Survival of mammalian embryos after storage at sub-zero temperatures was first achieved in the mouse in which all the preimplantation stages were shown to survive storage at \m=-\196\s=deg\C when suitably cooled and thawed (Whittingham, Leibo & Mazur, 1972;Wilmut, 1972;Whittingham, 1974a (Carworth-Europe). The day on which spermatozoa or a coital plug was found in the vagina was taken as Day 1 of pregnancy. Two-cell and four-cell embryos were flushed from the oviducts on the morning of Day 2 and Day 3, respectively, and eight-cell embryos were flushed from the oviduct and anterior portion of the uterine horn on Day 4 of pregnancy. Between ten and eighteen embryos were collected from each female. The embryos were collected in a standard culture medium for mouse embryos containing 94-59 mM-NaCl, 4-78 mM-KCl, 1-71 mM-CaCl2, 1-19 mM-MgS04.7H20, 1-19 mM-KH2P04, 25-07 raMNaHC03, 23-28 mM-Na lactate, 0-33 mM-Na pyruvate, 5-56 mM-glucose, 4 mg bovine serum albumin/ml, 100 Units penicillin G (potassium salt)/ml and 50 µg streptomycin sulphate/ml (Whittingham, 1971). They were washed in two changes of medium and transferred to 0-1 ml phosphate-buffered medium (PB1 : Whittingham & Wales, 1969) contained in a glass culture tube (12 x 75 mm). The embryos were equilibrated with 1-5 M-dimethylsulphoxide (DMSO) at 0°C for 15 min by the addition of a further 0-1 ml PB1 containing 3 m-DMSO. The procedures for freezing and thawing were similar to those used for mouse embryos and have been described elsewhere (Whittingham et al.,

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The Culture of Preimplantation Rat Embryos and the Production of Allophenic Rats

Cited by 62 publications

References 7 publications

Development of Mongolian Gerbil 1-cell Embryos to Blastocysts in Co-culture with Oviductal Cells in TCM 199 Supplemented with Pyruvate, Lactate and Fetal Calf Serum.

Development of Mongolian Gerbil 1-cell Embryos to Blastocysts in Co-culture with Oviductal Cells in TCM 199 Supplemented with Pyruvate, Lactate and Fetal Calf Serum.

A molecular basis for developmental plasticity in early mammalian embryos

Survival of Rat Embryos After Freezing and Thawing

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