scholarly journals Microtubular spindle dynamics and chromosome complements from somatic cell nuclei haploidization in mature mouse oocytes and developmental potential of the derived embryos

2004 ◽  
Vol 19 (5) ◽  
pp. 1181-1188 ◽  
Author(s):  
S.-U. Chen
Keyword(s):  
Somatic Cell ◽  
Cell Nuclei ◽  
Developmental Potential ◽  
Mouse Oocytes ◽  
Spindle Dynamics ◽  
Mature Mouse

Developmental potential of human oocytes reconstructed by transferring somatic cell nuclei into polyspermic zygote cytoplasm

2009 ◽  
Vol 382 (1) ◽  
pp. 119-123 ◽  
Author(s):  
Yong Fan ◽  
Xinjie Chen ◽  
Yumei Luo ◽  
Xiaolin Chen ◽  
Shaoying Li ◽  
...  
Keyword(s):  
Somatic Cell ◽  
Cell Nuclei ◽  
Developmental Potential ◽  
Human Oocytes

79 PSEUDOPHYSIOLOGICAL TRANSCOMPLEMENTARY ACTIVATION OF GOAT OOCYTES RECEIVING ADULT EAR CUTANEOUS FIBROBLAST CELL NUCLEI

2010 ◽  
Vol 22 (1) ◽  
pp. 198
Author(s):  
M. Skrzyszowska ◽  
M. Samiec
Keyword(s):  
Somatic Cell ◽  
Plasma Membranes ◽  
Dermal Fibroblast ◽  
Fibroblast Cell ◽  
Vero Cells ◽  
Atmospheric Conditions ◽  
Cell Nuclei ◽  
Developmental Potential ◽  
Water Saturated

The aim of the study was to determine the in vitro developmental potential of caprine cloned embryos following pseudophysiological (transcytoplasmic) transcomplementary activation of oocytes reconstructed with ear skin-derived fibroblast cell nuclei. The source of nuclear recipient cells were IVM doe oocytes. The reconstruction of the previously enucleated oocytes (i.e. ooplasts) was performed by microinjection of either the somatic cell-derived karyoplasts or intact whole tiny nuclear donor cells directly into the cytoplasm. The reconstructed oocytes were incubated in Upgraded B2 INRA medium for 30 min to 1 h before their pseudophysiological activation. The activation was achieved by electrofusion of clonal cybrids with the allogeneic cytoplasts isolated from caprine IVF-created zygotes, which led to the formation of triple allocytoplasmic hybrids (allocybrids). These originate from 3 sources: (1) homologous whole nuclear donor fibroblast cells or their karyoplasts; (2) enucleated oocytes (ooplasts), and (3) zygote-derived cytoplasts. Single zygote-descended cytoplasts (the so-called zygoplasts) were inserted into the perivitelline space of previously reconstituted oocytes. The resulting zygoplast-clonal cybrid couplets were subsequently subjected to electrofusion, which was induced by application of a single DC pulse of 2.4 kV cm-1 for 15 μs. The electrofusion of zygoplast and reconstructed oocyte plasma membranes occurred in an isotonic dielectric solution deprived of Ca2+ ions. The transcytoplasmically activated clonal cybrids were cultured in vitro in Upgraded B2 INRA medium for 48 h at 38.5°C in a 100% water-saturated atmosphere of 5% CO2 and 95% air. Afterward, cleaved embryos were co-cultured with Vero cells in medium supplemented with 10% fetal bovine serum for an additional 96 to 144 h up to morula and blastocyst stages under the same thermal and atmospheric conditions. A total of 53/78 (67.9%) oocytes reconstructed with fibroblast cell nuclei were successfully fused with zygoplasts. From among 53 cultured cloned embryos, 34 (64.2%) cleaved. The rates of embryos that reached the morula and blastocyst stages were 21/53 (39.6%) and 11/53 (20.8%), respectively. In conclusion, the relatively high percentages of morulae and blastocysts were noticed among in vitro-cultured caprine cloned embryos produced by the strategy of pseudophysiological transcytoplasmic activation of oocytes reconstructed with adult dermal fibroblast cell nuclei. Therefore, the use of cytoplasmic components originating from zygotes as the stimuli for activation of nuclear-transferred oocytes appeared to be an effective procedure in the generation of goat blastocysts by somatic cell cloning.

Effect of delayed enucleation on the developmental potential of nuclear-transferred oocytes receiving adult and fetal fibroblast cells

Zygote ◽  
2002 ◽  
Vol 10 (3) ◽  
pp. 217-222 ◽  
Author(s):  
Xi Jun Yin ◽  
Yoko Kato ◽  
Yukio Tsunoda
Keyword(s):  
Somatic Cell ◽  
Cell Number ◽  
Heart Beat ◽  
Fibroblast Cells ◽  
Cell Nuclei ◽  
Average Cell ◽  
Developmental Potential ◽  
Membrane Protrusions ◽  
Implantation Sites

To enhance the probability of reprogramming somatic cell nuclei, fibroblast cells from an adult male rabbit and a 12-day-old fetus were fused with oocytes at the second metaphase. The chromosomes of recipient oocytes were then removed by treatment with demecolcine for 1 or 2 h after fusion. Demecolcine treatment of fused oocytes induced membrane protrusions that contained all the maternal chromosomes, thus making it possible to remove the chromosomes. The potential of nuclear-transferred oocytes to develop into blastocysts was high (48% and 59%) and the average cell number of the blastocysts was large (149 and 159) 96 h after in vitro culture. The proportions of nuclear-transferred oocytes enucleated 1 h after fusion and implanted after transfer to pseudopregnant recipients were relatively high (2.8% and 4.9%) compared with our previous reports (1.7%: Yin et al., 2000; 0.6% and 1.0%: Yin et al., 2002a) where donor cells were fused with previously enucleated oocytes. Of 34 adult somatic cell implantation sites, 6 had fetuses on day 12 or 14 of pregnancy, but none of the fetuses had a heart beat or developed to term. None of the nuclear-transferred oocytes whose chromosomes were removed 2 h after demecolcine treatment implanted after transfer to recipients. The possible reasons why the high-quality nuclear-transferred oocytes did not develop to term are discussed.

Fusion with Activated Mouse Oocytes Modulates the Transcriptional Activity of Introduced Somatic Cell Nuclei

1996 ◽  
Vol 225 (1) ◽  
pp. 93-101 ◽  
Author(s):  
Ewa Borsuk ◽  
Maria S. Szöllösi ◽  
Didier Besombes ◽  
Pascale Debey
Keyword(s):  
Somatic Cell ◽  
Transcriptional Activity ◽  
Cell Nuclei ◽  
Mouse Oocytes

The effect of the time interval between injection and parthenogenetic activation on the spindle formation and the in vitro developmental potential of somatic cell nuclear-transferred rat oocytes

Zygote ◽  
2009 ◽  
Vol 18 (1) ◽  
pp. 9-15 ◽  
Author(s):  
Shigetoshi Mizumoto ◽  
Yoko Kato ◽  
Yukio Tsunoda
Keyword(s):  
Somatic Cell ◽  
Time Interval ◽  
Cell Nuclei ◽  
Spindle Formation ◽  
Developmental Potential ◽  
Developmental Rates ◽  
Culture Duration ◽  
Oocyte Cytoplasm ◽  
Preincubation Time

SummaryWe examined the optimal conditions for somatic cell nuclear transfer (SCNT) in the rat. First, we examined the effect of preincubation time before activation on SCNT rat oocytes produced in the presence of MG132 with regard to spindle formation and the potential to develop into blastocysts. The spindles of SCNT oocytes continued to elongate with an increase in the culture duration and, in approximately half of oocytes, the chromosomes were distributed along the spindles at 120 min after incubation. Such abnormal spindle formation in SCNT oocytes is a possible reason for the low developmental potential of SCNT rat oocytes. To inhibit the formation of abnormal spindle formation, we examined secondly the developmental potential of rat SCNT oocytes that had been preincubated with nocodazole and demecolcine instead of MG132. The developmental rates in SCNT oocytes, however, were decreased. For successful rat somatic cell cloning, two steps might be required: (1) to culture the somatic cell nuclei for a sufficient time in MII oocyte cytoplasm to enhance nuclear reprogramming; and (2) to induce normal spindle formation with normal chromosomal construction.

scholarly journals Developmental ability of cloned embryos from neural stem cells

Reproduction ◽  
2006 ◽  
Vol 132 (6) ◽  
pp. 849-857 ◽  
Author(s):  
Eiji Mizutani ◽  
Hiroshi Ohta ◽  
Satoshi Kishigami ◽  
Nguyen Van Thuan ◽  
Takafusa Hikichi ◽  
...  
Keyword(s):  
Somatic Cell ◽  
Success Rate ◽  
Sertoli Cells ◽  
Donor Cell ◽  
Cumulus Cells ◽  
Cloning Efficiency ◽  
Cell Nuclei ◽  
Developmental Potential ◽  
Undifferentiated State

The success rate is generally higher when cloning mice from embryonic stem (ES) cell nuclei than from somatic cell nuclei, suggesting that the embryonic nature or the undifferentiated state of the donor cell increases cloning efficiency. We assessed the developmental ability of cloned embryos derived from cultured neural stem cell (NSC) nuclei and compared the success rate with that of embryos cloned from other donor cells such as differentiated NSCs, cumulus cells, Sertoli cells and ES cells in the mouse. The transfer of two-cell cloned embryos derived from cultured NSC nuclei into surrogate mothers produced five live cloned mice. However, the success rate (0.5%) was higher in embryos cloned from cultured NSC nuclei than from differentiated NSCs (0%), but lower than that obtained by cloning mice from other cell nuclei (2.2–3.5%). Although thein vitrodevelopmental potential to the two-cell stage of the cloned embryos derived from NSC nuclei (73%) was similar to that of the cloned embryos derived from other somatic cell nuclei (e.g., 85% in Sertoli cells and 75% in cumulus cells), the developmental rate to the morula–blastocyst stage was only 7%. This rate is remarkably lower than that produced from other somatic cells (e.g., 50% in Sertoli cells and 54% in cumulus cells). These results indicate that the undifferentiated state of neural cells does not enhance the cloning efficiency in mice and that the arrest point forin vitrodevelopment of cloned embryos depends on the donor cell type.

Reconstruction of Mouse Oocytes With Somatic Cell Nuclei Produces Abnormal Meiotic Spindles

2005 ◽  
Vol 84 ◽  
pp. S387
Author(s):  
Q. Zhan ◽  
G. Zhuang ◽  
R. Li ◽  
Z. Cai ◽  
M. Kuwayama ◽  
...  
Keyword(s):  
Somatic Cell ◽  
Cell Nuclei ◽  
Mouse Oocytes ◽  
Meiotic Spindles

scholarly journals Introduction of Mouse Embryonic Fibroblasts into Early Embryos Causes Reprogramming and (Con)fusion

Cells ◽  
2021 ◽  
Vol 10 (4) ◽  
pp. 772
Author(s):  
Pierre Savatier
Keyword(s):  
Somatic Cell ◽  
Cell Nuclei ◽  
Mouse Embryonic Fibroblasts ◽  
Embryonic Fibroblasts ◽  
Early Embryos

The reprogramming of somatic cell nuclei to achieve pluripotency is one of the most important biological discoveries of the last few decades [...]

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