Birth of Cloned Calves Derived from Cultured Oviductal Epithelial Cells of a Dairy Cow

Goto, Yuji; Kaneyama, Kanako; Kobayashi, Sayo; Imai, Kyoichi; Shin-Noh, Masayuki; Tsujino, Takashi; Nakano, Tatsuya; Matsuda, Shuichi; Nakane, S.; Kojima, Toshiyuki

doi:10.2508/chikusan.70.243

Cited by 21 publications

(24 citation statements)

References 7 publications

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“…cattle from various somatic cell types derived from donor animals of different ages [15]. Moreover, we have generated offspring from these cloned cows by artificial insemination with conventionally processed spermatozoa from noncloned bulls [16].…”

Section: Telomere Lengths In Clone Offspringmentioning

confidence: 99%

Cloned Cows with Short Telomeres Deliver Healthy Offspring with Normal-length Telomeres

Miyashita

Kubo

Yonai

et al. 2011

Journal of Reproduction and Development

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Abstract. Dolly, the first mammal cloned from a somatic cell, had shorter telomeres than age-matched controls and died at an early age because of disease. To investigate longevity and lifetime performance in cloned animals, we produced cloned cows with short telomeres using oviductal epithelial cells as donor cells. At 5 years of age, despite the presence of short telomeres, all cloned cows delivered multiple healthy offspring following artificial insemination with conventionally processed spermatozoa from noncloned bulls, and their milk production was comparable to that of donor cows. Moreover, this study revealed that the offspring had normal-length telomeres in their leukocytes and major organs. Thus, cloned animals have normal functional germ lines, and therefore germ line function can completely restore telomere lengths in clone gametes by telomerase activity, resulting in healthy offspring with normal-length telomeres. Key words: Aging, Cattle, Mammalian cloning, Germ line, Telomere (J. Reprod. Dev. 57: [636][637][638][639][640][641][642] 2011) n mammals, most somatic cells undergo a finite number of cell divisions and ultimately enter a nondividing state known as replicative senescence [1] as a consequence of structural changes that take place in chromosomes over time. Chromosomes terminate in a nucleoprotein complex termed a telomere, which consists of repetitive sequences of G-rich noncoding DNA (TTAGGG)n and specific proteins. Telomeres are attached to the nuclear matrix and protect chromosome ends from degradation, fusion and recombination [2]. However, conventional DNA polymerases cannot replicate the extreme 5' ends of chromosomes because removal of the most terminal RNA primer in the lagging strand leaves a small region of uncopied DNA; therefore, telomeres are incrementally eroded with each cell division [3]. On the basis of this mechanism, it has been proposed that progressive telomere shortening during each cell division eventually yields a threshold telomere length beyond which additional normal cell divisions are not possible. Thus, telomere length acts as a mitotic clock that accounts for the limited lifespan of cells [4].In recent years, animal production by somatic cell nucleus transfer (SCNT) has offered a range of opportunities in basic and applied research, agriculture, genetic conservation and human medicine [5]. Somatic cell-cloned animals are produced using donor cells that have variably aged in vivo and in vitro, raising interesting questions about possible foreshortening effects on the lifespan and reproductive potential of these animals. In some cases, particularly with respect to cows and sheep, there is also the question of potential reductions in overall milk production. In this context, the telomere lengths in Dolly, the first mammal produced by SCNT, have been reported to be significantly shorter than those in age-matched controls, and this was consistent with the age of the donor mammary epithelial cell culture, which was derived from a 6-year-old donor sheep [6]. Dolly di...

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Section: Telomere Lengths In Clone Offspringmentioning

confidence: 99%

Cloned Cows with Short Telomeres Deliver Healthy Offspring with Normal-length Telomeres

Miyashita

Kubo

Yonai

et al. 2011

Journal of Reproduction and Development

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“…Briefly, bovine ovaries were collected at a slaughterhouse and stored in physiological saline at 20 C for 20 h. Oocytes were aspirated from follicles (2-5 mm in diameter) and cultured for 17-20 h in TCM199 containing 5% calf serum (CS), 5 mM Hepes, penicillin and streptomycin under 5% CO2 in air at 38.5 C. The cumulus cells were then removed by vortexing in 0.1% hyaluronidase, and the matured oocytes were collected for use as the recipient cytoplasm. Enucleation was performed using the zona pellucida cutting and nucleocytoplasm pushing methods [23]. …”

mentioning

confidence: 99%

Interspecies Nuclear Transfer Embryos Reconstructed from Cat Somatic Cells and Bovine Ooplasm

et al. 2008

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Abstract. This study was conducted to investigate the developmental capacity of domestic cat-bovine reconstructed embryos via interspecies somatic cell nuclear transfer (iSCNT) and to observe the mitochondrial DNA (mtDNA) content of the iSCNT embryos. The iSCNT embryos were generated using mixed-breed domestic cat fibroblasts as donor cells and enucleated bovine oocytes as the recipient cytoplasm. When the developmental capacities of iSCNT embryos and parthenogenic bovine embryos were compared, there was no difference (P>0.05) in the rates of cleavage and development to the 8-cell stage (86.6 vs. 84.0% and 32.2 vs. 36.2%, respectively). However, in contrast to development of parthenogenic embryos to the morula and blastocyst stages, no iSCNT embryos (0/202) developed beyond the 8-cell stage. For mtDNA analysis, iSCNT embryos at the 1-cell, 2-cell, 4-cell and 8-cell stages were randomly selected. Both cat and bovine mtDNA quantification analysis were performed using quantitative PCR. The levels of both cat and bovine mtDNA in cat-bovine iSCNT embryos varied at each stage of development. The cat mtDNA concentration in the iSCNT embryos was stable from the 1-cell to 8-cell stages. The bovine mtDNA in the iSCNT embryos at the 8-cell stage was significantly lower than that at the 4-cell stage (P<0.05). No difference in the proportions of cat mtDNA in the iSCNT embryos was found in any of the observed developmental stages (1-through 8-cell stages). In conclusion, bovine cytoplasm supports domestic cat nucleus development through the 8-cell stage. The mtDNA genotype of domestic catbovine iSCNT embryos illustrates persistence of heteroplasmy, and the reduction in mtDNA content might reflect a developmental block at the 8-cell stage. Key words: Bovine oocyte, Cat, Interspecies somatic cell nuclear transfer (iSCNT), Mitochondrial DNA analysis (J. Reprod. Dev. 54: [142][143][144][145][146][147] 2008) nterspecies somatic cell nuclear transfer (iSCNT) involves transfer of donor cell nuclei from one species to the enucleated oocytes of another species. The type of nuclear transfer allows for derivation of embryos that carry valuable genes in a surrogate ooplasm from another mammalian species. This technology was established for many reasons including production of embryos from species from which oocytes are difficult to obtain or where their collection is under restricted control. In regard to reproductive aims, iSCNT has been applied to conservation of wildlife animals including gaur [1], mouflon [2], banteng [3] and African wildcats [4]. In regard to therapeutic or biomedical aims, patientspecific embryonic cell lines have been established using iSCNT [5,6]. Furthermore, iSCNT provides an opportunity for molecular tracking of nucleocytoplasmic interaction and the transmission of two cytoplasmic populations. This methodology influences embryonic development and survival post-implantation and provides an enabling technology to explore fundamental aspects of developmental biology in mammals.The survival of wildcats in Thail...

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“…In the previous studies on cattle somatic cell NT [8][9][10]17], no specified donor cell size was mentioned. In bovine embryonic cell lines cultured under non-serum-starvation conditions, it is generally accepted that small cells have divided more recently, and therefore are earlier in the cell cycle (G1 phase) [18].…”

Section: Discussionmentioning

confidence: 99%

Bovine Nuclear Transfer Using Cumulus Cells Derived from Serum-Starved and Confluent Cultures

Nour

Ikeda

Takahashi

2000

Journal of Reproduction and Development

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Abstract. Bovine cumulus cells after oocyte maturation were cultured under serum-starvation condition or grown to confluence. Their cell size, cell cycle phases, and developmental potential as donors for nuclear transfer (NT) were investigated. The majority of trypsinized cells of both cultures were found within the medium size range (15-20 µm). Flow cytometric analysis revealed that small-and medium-sized cells had significantly higher percentages of nuclei existing in the G0/G1 phase (95-98%) than large-sized cells (82%) regardless of the culture conditions. Serumstarved cells of different sizes (small, 9-14 µm; medium, 15-20 µm; large, 21-26 µm) had the same fusion rate, cleavage, development to blastocysts, and blastocyst cell number of NT embryos. NT embryos reconstituted from serum-starved and confluent medium-sized cells showed the same developmental rate. These results indicate that the most available medium-sized cells derived from both serum-starvation and confluent cultures can be used as a G0/G1-phase nuclei source. Key words: Bovine, Cell size, Cumulus cells, Nuclear transfer.(J. Reprod. Dev. 46: [85][86][87][88][89][90][91][92] 2000) than larger cells when serum-starved cultured porcine fetal fibroblasts were used as donor nuclei in the nuclear transfer (NT) procedure [2,3]. In view of this, one could imagine that the use of small donor cells would lead to higher embryonic development after nuclear transfer. However, small and large mouse cumulus cells that are naturally arrested in the G0 phase seldom support the development of reconstituted embryos beyond the 8-cell stage [4]. In this respect, no available study concurrently investigated the cell cycle phase of the donors as related to the cell size and the consequent development after nuclear transfer. The recent success in getting live offspring after somatic cell nuclear transfer using fetal fibroblasts [5,6], adult mammary gland cells [6], and cumulus cells [7,8] indicates that successful reprogramming of donor chromatin from differentiated cells may be related to serum starvation of donor cells which uccessful production of offspring derived after somatic cell nuclear transfer depends upon a variety of factors. The most important factor identified thus far is the appropriate cell cycle coordination of donor nuclei and recipient oocytes [1]. To condense normally and maintain correct ploidy, donor nuclei in the G0/G1 phase should be transferred to metaphase II (MII) arrested oocytes with a high level of maturation promoting factor (MPF). Another factor that may be of importance is the size of the donor cell. Recent flow cytometric study of porcine fetal fibroblasts [2] showed that as the cell size decreased, the percentages of cells existing in G0+G1 and G0 phases increased significantly. In addition, small cells were more likely to have nuclear formation

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Birth of Cloned Calves Derived from Cultured Oviductal Epithelial Cells of a Dairy Cow

Cited by 21 publications

References 7 publications

Cloned Cows with Short Telomeres Deliver Healthy Offspring with Normal-length Telomeres

Cloned Cows with Short Telomeres Deliver Healthy Offspring with Normal-length Telomeres

Interspecies Nuclear Transfer Embryos Reconstructed from Cat Somatic Cells and Bovine Ooplasm

Bovine Nuclear Transfer Using Cumulus Cells Derived from Serum-Starved and Confluent Cultures

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