The Culture of Fibroblasts From Diaphragm of Giant Panda

Han, Zhiming; Chen, Da‐Yuan; Li, Jinsong; Sun, Qing‐Yuan; Wang, Peng-Yang; Huang, Yang; Du, Jun

doi:10.1290/1071-2690(2001)037<0644:tcoffd>2.0.co;2

Cited by 16 publications

(10 citation statements)

References 2 publications

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“…The cumulus cells were cultured at 378C to reach confluence, and then passaged with routine methods. We obtained rabbit skin fibroblasts from a fetus at day 17 of gestation as described before (Han et al, 2001;Chen et al, 2004). Both porcine and rabbit cells at passages 3-10 were used as donors.…”

Section: Preparation Of Donor Cellsmentioning

confidence: 99%

Interspecies nuclear transfer reveals that demethylation of specific repetitive sequences is determined by recipient ooplasm but not by donor intrinsic property in cloned embryos

et al. 2006

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DNA methylation/demethylation of donor genomes in recipient ooplasm after nuclear transfer occurs in a species-specific way. In cloned rabbit and bovine embryos, repetitive sequences maintain the donor-type methylation status, but typical demethylation of repetitive sequences takes place in cloned porcine embryos. To clarify whether the demethylation is controlled by donor nucleus intrinsic property or by recipient ooplasm, we used interspecies somatic cell nuclear transfer (iSCNT) model to examine the methylation status of repetitive sequences in pig-to-rabbit and rabbit-to-pig interspecies embryos. We found that no demethylation of pig repetitive sequences was observed in pig-to-rabbit iSCNT embryos, while the examined rabbit repetitive sequence Rsat IIE was demethylated in rabbit-to-pig iSCNT embryos. These results indicate that demethylation of donor repetitive sequences is determined by ooplasm but not by donor intrinsic property and that ooplasm from different species have different capabilities to demethylate genes.

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Section: Preparation Of Donor Cellsmentioning

confidence: 99%

Interspecies nuclear transfer reveals that demethylation of specific repetitive sequences is determined by recipient ooplasm but not by donor intrinsic property in cloned embryos

et al. 2006

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“…The method of cell culture and assessment has been described previously (Han et al, 2001). Briefly, ear tissue specimens derived from a 6-year-old female adult Asian yellow goat were finely chopped into pieces measuring about 1 mm 2 and enzymatically dissociated using 0.25% trypsin and 1 mM EDTA (Gibco BRL, Grand Island, NY) in PBS (Gibco BRL) for 12 hr at 48C and then for 30 min at 378C.…”

Section: Preparation Of Donor Cellsmentioning

confidence: 99%

Cloning of Asian yellow goat (C. hircus) by somatic cell nuclear transfer: Telophase enucleation combined with whole cell intracytoplasmic injection

Chen

Jiang

Zhao

et al. 2006

Molecular Reproduction Devel

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Our and other previous studies have shown that telophase enucleation is an efficient method for preparing recipient cytoplasts in nuclear transfer. Conventional methods of somatic cell nuclear transfer either by electro-fusion or direct nucleus injection have very low efficiency in animal somatic cell cloning. To simplify the manipulation procedure and increase the efficiency of somatic cell nuclear transfer, this study was designed to study in vitro and in vivo development of Asian yellow goat cloned embryos reconstructed by direct whole cell intracytoplasmic injection (WCICI) into in vitro matured oocytes enucleated at telophase II stage. Our results demonstrated that the rates of cleavage and blastocyst development of embryos reconstructed by WCICI were slightly higher than in conventional subzonal injection (SUZI) group, but no statistic difference (P > 0.05) existed between these two methods. However, the percentage of successful embryonic reconstruction in WCICI group was significantly higher than that in SUZI group (P < 0.05). After embryo transfer at 4-cell stage, the foster in both groups gave birth to offspring. Therefore, the present study suggests that the telophase ooplasm could properly reprogram the genome of somatic cells, produce Asian yellow goat cloned embryos and viable kids, and whole cell intracytoplasmic injection is an efficient protocol for goat somatic cell nuclear transfer.

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“…Primary cell culture was performed with the same method as that described previously (Han et al 2001). Fibroblasts at passages 4-10 were used as nuclear donors.…”

Section: Reconstruction Of Macaque -Rabbit Cloned Embryosmentioning

confidence: 99%

Quantitative analysis of mitochondrial DNAs in macaque embryos reprogrammed by rabbit oocytes

Yang

Kou²,

Wang³

et al. 2004

Reproduction

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In cloned animals where somatic cell nuclei and oocytes are from the same or closely related species, the mitochondrial DNA (mtDNA) of the oocyte is dominantly inherited. However, in nuclear transfer (NT) embryos where nuclear donor and oocyte are from two distantly related species, the distribution of the mtDNA species is not known. Here we determined the levels of macaque and rabbit mtDNAs in macaque embryos reprogrammed by rabbit oocytes. Quantification using a realtime PCR method showed that both macaque and rabbit mtDNAs coexist in NT embryos at all preimplantation stages, with maternal mtDNA being dominant. Single NT embryos at the 1-cell stage immediately after fusion contained 2.6 3 10 4 copies of macaque mtDNA and 1.3 3 10 6 copies of rabbit mtDNA. Copy numbers of both mtDNA species did not change significantly from the 1-cell to the morula stages. In the single blastocyst, however, the number of rabbit mtDNA increased dramatically while macaque mtDNA decreased. The ratio of nuclear donor mtDNA to oocyte mtDNA dropped sharply from 2% at the 1-cell stage to 0.011% at the blastocyst stage. These results suggest that maternal mtDNA replicates after the morula stage.

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The Culture of Fibroblasts From Diaphragm of Giant Panda

Cited by 16 publications

References 2 publications

Interspecies nuclear transfer reveals that demethylation of specific repetitive sequences is determined by recipient ooplasm but not by donor intrinsic property in cloned embryos

Interspecies nuclear transfer reveals that demethylation of specific repetitive sequences is determined by recipient ooplasm but not by donor intrinsic property in cloned embryos

Cloning of Asian yellow goat (C. hircus) by somatic cell nuclear transfer: Telophase enucleation combined with whole cell intracytoplasmic injection

Quantitative analysis of mitochondrial DNAs in macaque embryos reprogrammed by rabbit oocytes

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