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J Vet Sci. 2007 Dec;8(4):415-421 |
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Autologous somatic cell nuclear transfer in pigs using recipient oocytes and donor cells from the same animal
Eunsong Lee1,*, Kilyoung Song2 |
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1School of Veterinary Medicine and Institute of Veterinary Science, Kangwon National University, Chunchon 200-701, Korea
2College of Veterinary Medicine, Seoul National University, Seoul 151-742, Korea
* eslee@kangwon.ac.kr |
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The objective of the present study was to examine the
feasibility of the production of autologous porcine somatic
cell nuclear transfer (SCNT) blastocysts using oocytes and
donor cells from slaughtered ovaries. Therefore, we attempted
to optimize autologous SCNT by examining the
effects of electrical fusion conditions and donor cell type
on cell fusion and the development of SCNT embryos.
Four types of donor cells were used: 1) denuded cumulus
cells (DCCs) collected from in vitro-matured (IVM) oocytes;
2) cumulus cells collected from oocytes after 22 h of
IVM and cultured for 18 h (CCCs); 3) follicular cells obtained
from follicular contents and cultured for 40 h
(CFCs); and 4) adult skin fibroblasts. The DCCs showed a
significantly (p £¼ 0.01) lower rate of fusion than the CCCs
when two pulses of 170 V/mm DC were applied for 50 ¥ìsec
(19 ¡¾ 2% vs. 77 ¡¾ 3%). The rate of DCC fusion with oocytes
was increased by the application of two DC pulses of
190 V/mm for 30 ¥ìsec, although this was still lower than
the rate of fusion in the CCCs (33 ¡¾ 1% vs. 80 ¡¾ 2%). The
rates of cleavage (57 ¡¾ 5%) and blastocyst formation (1 ¡¾
1%) in the DCC-derived embryos did not differ from
those (55 ¡¾ 6% and 3 ¡¾ 1%, respectively) in the CCC-derived
SCNT embryos. Autologous SCNT embryos derived
from CFCs (5 ¡¾ 2%) showed higher levels of blastocyst
formation (p £¼ 0.01) than CCC-derived autologous SCNT
embryos (1 ¡¾ 0%). In conclusion, the results of the present
study show that culturing cumulus and follicular cells before
SCNT enhances cell fusion with oocytes and that
CFCs are superior to CCCs in the production of higher
numbers of autologous SCNT blastocysts.
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