Production of Transgenic Chimera Rabbit Fetuses Using Somatic Cell Nuclear Transfer

Matsuda, Junichi; Takahashi, Seiya; Ohkoshi, Katsuhiro; Kaminaka, Kazuyoshi; Kaminaka, Sara; Nozaki, Chikateru; Maeda, Hiroaki; Tokunaga, Tomoyuki

doi:10.1089/153623002753632002

Cited by 15 publications

(15 citation statements)

References 39 publications

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“…This method was also used to support the development ability of cloned embryos in rabbits by forming a chimera with normal embryos [22]. When applying this method to pigs, our data will be significant in regulating sex differentiation and germ-line chimerism in chimeric embryos.…”

Section: Discussionmentioning

confidence: 95%

Sex Differentiation and Germ Cell Production in Chimeric Pigs Produced by Inner Cell Mass Injection into Blastocysts

Nagashima¹,

Giannakis²,

Ashman³

et al. 2004

Biology of Reproduction

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This study aimed at collecting background knowledge for chimeric pig production. We analyzed the genetic sex of the chimeric pigs in relation to phenotypic sex as well as to functional germ cell formation. Chimeric pigs were produced by injecting Day 6 or Day 7 inner cell mass (ICM) cells into Day 6 blastocysts. Approximately 20% of the piglets born from the injected blastocysts showed overt coat color chimerism regardless of the embryonic stage of donor cells. The male:female sex ratio was 7:2 and 6:1 in the chimeras derived from Day 6 and Day 7 ICM cells, respectively, showing an obvious bias toward males. When XX donor cells were injected into XY blastocysts at the same embryonic stage, the phenotypic sex of the resulting chimera was male with no germ-line cells formed from the donor cell lineage. On the other hand, when the donor was XY and the recipient blastocyst was XX, the phenotypic sex of the chimera was male, and germ-line cells were derived only from the donor cells. The combination of XY donor cells and XY blastocysts produced some chimeras in which the donor cell lineage did not contribute to germ-line formation even when it appeared in coat color. When the embryonic stage of the donor was advanced by 1 day in the XY-XY combination, 100% of the germ-line cells of the chimeras were derived from the donor cell lineage. These data showed that characteristics of sex differentiation and germ cell formation in chimeric pigs are similar to those in chimeric mice.

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Section: Discussionmentioning

confidence: 95%

Sex Differentiation and Germ Cell Production in Chimeric Pigs Produced by Inner Cell Mass Injection into Blastocysts

Nagashima¹,

Giannakis²,

Ashman³

et al. 2004

Biology of Reproduction

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“…Therefore, it is possible that the parthenogenetic embryos could implant in the endometrium more efficiently and trigger a good pregnant status, which might be a better receptive status for the SCNT embryos to implant and to maintain pregnancy. Previous reports had shown that aggregation of blastomeres of cloned embryos with fertilized ones improved the development of the resulted embryos and achieved live chimera (Matsuda et al, 2002;Skrzyszowska et al, 2006); also, the cloned embryos showed better term development when they were combined with one or two parthenogenetic blastomeres rather than with the regular ones .…”

Section: Tsa Treatment and Pa Embryo Co-transfer In Rabbit Scnt 205mentioning

confidence: 86%

Live Birth of Somatic Cell-Cloned Rabbits following Trichostatin A Treatment and Cotransfer of Parthenogenetic Embryos

Meng

Polgár

et al. 2009

Cloning and Stem Cells

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Somatic cell nuclear transfer (SCNT) efficiency is still low in rabbit. Previous studies indicated that trichostatin A (TSA) treatment could improve cloning efficiency and term development in the mouse, and cotransfer of parthenogenetic (PA) embryos benefited the pregnancy of cloned embryos in porcine and the mouse. In this study we investigated the effect of TSA treatment on the term development of the SCNT rabbit embryos, and the possibility of the pregnancy maintenance of clones by cotransfer of PA embryos. The SCNT embryos were produced by fusing cumulus cells with enucleated cytoplasts before activation by electrical stimulation, and Dimethylaminopurine (6-DMAP) and Cyclohexamide (CHX) treatments. They were cultured in EBSS-complete medium regardless of their treatment with or without TSA. In vitro developmental data showed no differences in the cleavage and the blastocyst rates, and the blastocyst cell number between the TSA-treated and the untreated SCNT embryos. Two of the six recipients became pregnant after the embryo transfer (ET) in the TSA-treated group, and one pregnant female delivered seven live and three stillborn pups. The death of all live pups occurred within an hour to 19 days. Four of the seven recipients became pregnant in the TSA-untreated group. Three of them gave birth to six live and eight stillborn pups. Four pups of the TSA-untreated group have grown into adulthood, and three of them produced progeny. Cotransfer of three to four PA embryos with 26-32 SCNT embryos to the same recipient resulted in pregnancy and birth rates statistically no different compared to the control SCNT ET group. In conclusion, our results indicate that TSA treatment has a limited effect on the in vitro development of the SCNT embryos; furthermore, both the TSA-treated and the untreated clones can develop to term in rabbits, but none of the offspring from TSA-treated embryos survived to adulthood in our experiment.

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“…The rabbit not only is anatomically relevant to the human but also has a short gestation period (28–31 days), is relatively low maintenance and is easy to handle. Whereas SCNT and animal cloning has become routine (Li et al 2009; Matsuda et al 2002), rabbit cloning is considered difficult (Yang et al 2007). To date, only four reported cases worldwide have achieved success in SCNT in the rabbit.…”

Section: Discussionmentioning

confidence: 99%

“…Both the phylogenetic nature of the rabbit and its physiology and anatomy make the rabbit a relevant model for the biochemical, molecular and physiological characterization of CF pathology and for the development of CF therapies (Vuillaumier et al 1997; Zeitlin et al 1992). Since no viable embryonic stem (ES) cell systems exist that have been established for the development of transgenic animals and germline transfer outside of the mouse, the generation of large animal models is dependent on cloning technology and somatic cell nuclear transfer (SCNT; Challah-Jacques et al 2003; Kasinathan et al 2001; Matsuda et al 2002). …”

Section: Introductionmentioning

confidence: 99%

Generation of SV40-transformed rabbit tracheal-epithelial-cell-derived blastocyst by somatic cell nuclear transfer

Semir

Maurisse

et al. 2012

Cell Tissue Res

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The prospect of developing large animal models for the study of inherited diseases, such as cystic fibrosis (CF), through somatic cell nuclear transfer (SCNT) has opened up new opportunities for enhancing our understanding of disease pathology and for identifying new therapies. Thus, the development of species-specific in vitro cell systems that will provide broader insight into organ- and cell-type-specific functions relevant to the pathology of the disease is crucial. Studies have been undertaken to establish transformed rabbit airway epithelial cell lines that display differentiated features characteristic of the primary airway epithelium. This study describes the successful establishment and characterization of two SV40-transformed rabbit tracheal epithelial cell lines. These cell lines, 5RTEo- and 9RTEo-, express the CF transmembrane conductance regulator (CFTR) gene, retain epithelial-specific differentiated morphology and show CFTR-based cAMP-dependent Cl− ion transport across the apical membrane of a confluent monolayer. Immunocytochemical analysis indicates the presence of airway cytokeratins and tight-junction proteins in the 9RTEo- cell line after multiple generations. However, the tight junctions appear to diminish in their efficacy in both cell lines after at least 100 generations. Initial SCNT studies with the 9RTEo- cells have revealed that SV40-transformed rabbit airway epithelial donor cells can be used to generate blastocysts. These cell systems provide valuable models for studying the developmental and metabolic modulation of CFTR gene expression and rabbit airway epithelial cell biology.

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Production of Transgenic Chimera Rabbit Fetuses Using Somatic Cell Nuclear Transfer

Cited by 15 publications

References 39 publications

Sex Differentiation and Germ Cell Production in Chimeric Pigs Produced by Inner Cell Mass Injection into Blastocysts

Sex Differentiation and Germ Cell Production in Chimeric Pigs Produced by Inner Cell Mass Injection into Blastocysts

Live Birth of Somatic Cell-Cloned Rabbits following Trichostatin A Treatment and Cotransfer of Parthenogenetic Embryos

Generation of SV40-transformed rabbit tracheal-epithelial-cell-derived blastocyst by somatic cell nuclear transfer

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