scholarly journals Production of Blastocysts after Intergeneric Nuclear Transfer of Goral (Naemorhedus goral) Somatic Cells into Bovine Oocytes

2006 ◽  
Vol 68 (11) ◽  
pp. 1167-1171 ◽  
Author(s):  
Byoung-Chol OH ◽  
Jung-Tae KIM ◽  
Nam-Shik SHIN ◽  
Soo-Whan KWON ◽  
Sung-Keun KANG ◽  
...  
Keyword(s):  
Nuclear Transfer ◽  
Somatic Cells ◽  
Bovine Oocytes

44 NUCLEAR TRANSFER IN CATTLE USING SOMATIC CELLS FROM FROZEN TESTICLES WITHOUT CRYOPROTECTANTS

2007 ◽  
Vol 19 (1) ◽  
pp. 140 ◽  
Author(s):  
Y. Hoshino ◽  
N. Kobayashi ◽  
N. Hayashi ◽  
T. Matsuhashi ◽  
K. Saeki ◽  
...  
Keyword(s):  
Nuclear Transfer ◽  
Culture Medium ◽  
Somatic Cells ◽  
Frozen Storage ◽  
Cell Number ◽  
Fetal Skin ◽  
Nylon Mesh ◽  
Viable Cells ◽  
Donor Cells ◽  
Bovine Oocytes

Obtaining somatic cells from preserved organs or tissues is useful for the conservation and regeneration of genetic resources by nuclear transfer (NT). Bovine cells for NT have been obtained from cooled carcasses stored at 0�C for several days (Arat et al. 2005 Reprod. Fert. Dev. 17, 164 abst) and from fetal skin tissue cryopreserved with DMSO (Fahrudin et al. 2001 J. Vet. Med. Sci. 63, 1151–1154). However, frozen storage of organs or tissues without cryoprotectants was considered to be quite inappropriate for obtaining viable cells. We report here that viable donor cells for NT were obtained from bovine testicles after frozen storage without cryoprotectants. In the first experiment, we investigated whether viable cells can be recovered from frozen testicles castrated from Japanese Black bulls. The testicles were frozen at -80�C in a freezer for several days; then some were stored in liquid nitrogen for 10 months without cryoprotectants. Before thawing, the testicles were divided into 3 pieces, caput epididymis, cauda epididymis, and testis. Each piece was then put in saline at 42�C for quick thawing. Thawed tissues were minced into 5-mm pieces and incubated at 39�C for 2 h in DMEM containing 0.1% collagenase and 0.2% dispase. After filtration through a 250-�m nylon mesh filter, the filtrates were centrifuged at 250 � 4g for 5 min. Then precipitates were resuspended with MF-start� primary culture medium (TM Cell Research Inc., Fukui, Japan) and incubated at 38.5�C under the atmosphere of 5% CO2 in air with high humidity. After 5 days of incubation, the medium was replaced and nonadherent debris was discarded. Viable cells were obtained from the caput epididymis. These cells actively proliferated and expanded. In the next experiment, to determine whether these cells can be used for NT, the cells were electrically fused with enucleated bovine oocytes. Bovine fibroblasts taken from unfrozen ear tissue were used as controls. The NT embryos were activated by Ca-ionophore treatment, followed by treatment with cycloheximide for 6 h, and then cultured in mSOF for 168 h. NT embryos reconstructed from testicle cells did not significantly differ from NT embryos made with control cells with regard to blastocyst rates (22.1% and 20.2%), cell number of blastocysts [130 � 43 and 121 � 43 (mean � SD)], and ICM ratio (21.1% and 22.6%), respectively (ANOVA). These results suggest that somatic cells derived from bovine frozen testicles can be used for nuclear transfer. Further studies are needed to examine whether viable cells can be obtained from other frozen organs or tissues. This study was partially supported by a grant from the Wakayama Prefecture Collaboration of Regional Entities for the Advancement of Technological Excellence, JST.

Blastocyst Development after Intergeneric Nuclear Transfer of Mountain Bongo Antelope Somatic Cells into Bovine Oocytes

2003 ◽  
Vol 5 (1) ◽  
pp. 25-33 ◽  
Author(s):  
Byeongchun Lee ◽  
Gemechu G. Wirtu ◽  
Philip Damiani ◽  
Earle Pope ◽  
Betsy L. Dresser ◽  
...  
Keyword(s):  
Nuclear Transfer ◽  
Somatic Cells ◽  
Blastocyst Development ◽  
Bovine Oocytes

Cloned blastocysts produced by nuclear transfer from somatic cells in cynomolgus monkeys (Macaca fascicularis)

Primates ◽  
2007 ◽  
Vol 48 (3) ◽  
pp. 232-240 ◽  
Author(s):  
Junko Okahara-Narita ◽  
Hideaki Tsuchiya ◽  
Tatsuyuki Takada ◽  
Ryuzo Torii
Keyword(s):  
Nuclear Transfer ◽  
Macaca Fascicularis ◽  
Somatic Cells ◽  
Cynomolgus Monkeys

Cloned rabbits produced by nuclear transfer from adult somatic cells

2002 ◽  
Vol 20 (4) ◽  
pp. 366-369 ◽  
Author(s):  
Patrick Chesné ◽  
Pierre G. Adenot ◽  
Céline Viglietta ◽  
Michel Baratte ◽  
Laurent Boulanger ◽  
...  
Keyword(s):  
Nuclear Transfer ◽  
Somatic Cells

Nuclear transplantation and the control of gene activity in animal development

1970 ◽  
Vol 176 (1044) ◽  
pp. 303-314 ◽  
Keyword(s):  
Nuclear Transfer ◽  
Somatic Cells ◽  
Gene Activity ◽  
Protein Fractions ◽  
Nuclear Transplantation ◽  
Animal Development ◽  
Cytoplasmic Proteins ◽  
Intracellular Movement ◽  
Selective Accumulation ◽  
Structure And Metabolism

The transplantation of nuclei from differentiated or determined somatic cells to enucleated frogs’ eggs consistently leads to a complete and clearly recognizable change of gene activity. Within 1 to 2 h of nuclear transfer, somatic nuclei have come to resemble in structure and metabolism the zygote nuclei of fertilized eggs. The change in gene activity therefore takes place very soon after nuclear transfer and results from an effect of egg cytoplasm. The induced change in gene activity is associated with a selective accumulation of cytoplasmic proteins in transplanted nuclei. Examples are given of various ways in which nuclear transplantation and microinjection can be used to elucidate the intracellular movement of proteins and the effect of known protein fractions on gene activity.

scholarly journals 57EMBRYO DEVELOPMENT FOLLOWING INTERSPECIES NUCLEAR TRANSFER OF AFRICAN BUFFALO (SYNCERUS CAFFER), BONTEBOK (DAMALISCUS DORCUS DORCUS) AND ELAND (TAUROTRAGUS ORYX) SOMATIC CELLS INTO BOVINE CYTOPLASTS

2004 ◽  
Vol 16 (2) ◽  
pp. 150 ◽  
Author(s):  
M. Matshikiza ◽  
P. Bartels ◽  
G. Vajta ◽  
F. Olivier ◽  
T. Spies ◽  
...  
Keyword(s):  
South Africa ◽  
Nuclear Transfer ◽  
Somatic Cells ◽  
Blastocyst Stage ◽  
Critically Endangered ◽  
African Buffalo ◽  
Syncerus Caffer ◽  
Cell Stage ◽  
Wildlife Species ◽  
Interspecies Nuclear Transfer

Wildlife conservation requires traditional as well as innovative conservation strategies in order to preserve gene and species diversity. Interspecies nuclear transfer has the potential to conserve genes from critically endangered wildlife species where few or no oocytes are available from the endangered species, and where representative cell lines have been established for the wildlife population while numbers were still abundant. The purpose of this study was to investigate the developmental ability of embryos reconstructed with transfer of somatic cells from the African buffalo (Syncerus caffer), bontebok (Damaliscus dorcus dorcus) and eland (Taurotragus oryx) to enucleated domestic cattle (Bos taurus) oocytes. Skin tissue from the three wildlife species were collected by surgically removing approx. 1.0×1.0cm ear skin notches from animals immobilized with a combination of etorphine hydrochloride (M99; South Africa) and azaperone (Stressnil, South Africa). The biopsies were placed into physiological saline and transported to the laboratory at 4°C within 2h, cleaned with chlorohexidine gluconate and sliced finely in Minimal Essential Medium supplemented with 10% fetal calf serum. The resultant tissue explants were treated as previously described (Baumgarten and Harley 1995 Comp. Biochem. Physiol. 110B, 37–46) and actively growing fibroblast cultures made available for the nuclear transfer process. Nuclear transfer was performed using the HMC technique (Vajta et al., 2003 Biol. Reprod. 68, 571–578) using slaughterhouse-derived bovine oocytes. Culture was performed in SOFaaci (Vajta et al., 2003 Biol. Reprod. 68, 571–578) medium supplemented with 5% cattle serum using WOWs (Vajta et al., Mol. Reprod. Dev. 50, 185–191). Two identical replicates were made with somatic cells of each species. After successful reconstruction, 57, 42 and 48 nuclear transferred and activated buffalo, bontebok and eland embryos were cultured, respectively. All except for 2 buffalo embryos cleaved; 22 (39%) developed to or over the 8-cell stage, and 2 (3.5%) of them to the blastocyst stage. All but 3 bontebok embryos cleaved, 17 (40%) developed to or over the 8-cell stage, but none of them reached the compacted morula or blastocyst stage. Sixteen (33%) of the eland embryos developed to or over the 8-cell stage with one (2%) reaching the blastocyst stage. In conclusion, buffalo, bontebok and eland embryos developed from reconstruction using their respective somatic cells combined with bovine cytoplasts, however, in vitro developmental ability to the blastocyst stage was limited. Additional basic research that establishes the regulative mechanisms involved with early preimplantation development together with optimising nuclear transfer techniques may have the potential to one day play a role in the conservation of critically endangered wildlife species.

scholarly journals Erratum: Production of gene-targeted sheep by nuclear transfer from cultured somatic cells

Nature ◽  
2000 ◽  
Vol 408 (6808) ◽  
pp. 120-120 ◽  
Author(s):  
K. J. McCreath ◽  
J. Howcroft ◽  
K. H. S. Campbell ◽  
A. Colman ◽  
A. E. Schnieke ◽  
...  
Keyword(s):  
Nuclear Transfer ◽  
Somatic Cells

The localization of LAP2β during pronuclear formation in bovine oocytes after fertilization or activation

Zygote ◽  
2006 ◽  
Vol 14 (2) ◽  
pp. 157-167 ◽  
Author(s):  
Mamiko Isaji ◽  
Hisataka Iwata ◽  
Hiroshi Harayama ◽  
Masashi Miyake
Keyword(s):  
Chromatin Condensation ◽  
Polar Body ◽  
Somatic Cells ◽  
Histone H3 ◽  
Cumulus Cells ◽  
Polar Bodies ◽  
Second Polar Body ◽  
Pronuclear Formation ◽  
Bovine Oocytes

SummaryWe have shown that the assembly of lamin-associated polypeptide (LAP) 2β was detected surrounding the chromatin mass around the time of extrusion of the second polar body (PB) in some fertilized oocytes, but not in most activated oocytes, by using A23187 and cycloheximide (CaA + CH). Here, we immunohistologically analysed the correlation between LAP2β assembly and chromatin condensation in fertilized and activated oocytes during the second meiosis. In bovine cumulus cells, the onset of LAP2β assembly was observed around anaphase chromosomes with strongly phosphorylated histone H3. No LAP2β assembled around the chromosomes in the first and second polar bodies and the alternative oocyte chromatin (oCh) if histone H3 was phosphorylated. Only histone H3 of oCh was completely dephosphorylated during the telophase II/G1 transition (Tel II/G1), and then LAP2β assembled around only the oCh without phosphorylated histone H3. In the oocytes activated by CaA + CH, LAP2β did not assemble around the condensed oCh during the Tel II/G1 transition, although their histone H3 dephosphorylation occurred rather rapidly compared with that of the fertilized oocytes. The patterns of histone H3 dephosphorylation and LAP2β assembly in oocytes activated by CaA alone showed greater similarity to those in fertilized oocytes than to those in oocytes activated by CaA + CH. These results show that LAP2β assembles around only oCh after complete dephosphorylation of histone H3 after fertilization and activation using CaA alone, and that the timing of histone H3 dephosphorylation and LAP2β assembly in these oocytes is different from that of somatic cells. The results also indicate that CH treatment inhibits LAP2β assembly around oCh but not histone H3 dephosphorylation.

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