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In vitro developmental potential of interspecies nuclear transfer embryos using bovine oocytes and mouse embryonic fibroblasts

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Authors
구본식
Advisor
이창규
Issue Date
2004
Publisher
서울대학교 대학원
Keywords
이종간 핵이식Interspecies nuclear transferBlock 배양액Block media항산화제Antioxidant유전자 발현Gene expression
Description
Thesis (master`s)--서울대학교 대학원 :농생명공학부,2004.
Abstract
Development of somatic cell nuclear transfer(NT) technology to produce clone animals opens a new
era in the field of reproductive biology and medicine, and the birth of clones was reported in
several species. However, the efficacy of this system still remains low and oocytes
reconstructed with somatic cells had lower developmental competence than embryos derived from in
vitro fertilization. In using bovine oocytes as a recipient, embryos have been obtained through
NT of somatic cells from several cell types. Although recipient oocytes are not derived from
the same species used for a donor, NT generates interspecies NT embryos. They might be used to
find answers of some questions in basic aspect of NT. Therefore, we investigated the development
potential of interspecies NT embryos using mouse embryonic fibroblasts and examine blastocysts
rates, antioxidants effect and gene expression.
To investigate the effect of bovine cytoplast on developmental block of interspecies NT
embryos, two groups of media were used. One group is consisted of mouse culture media with
normal (able to overcome developmental block) and block (induce 2-cell block). The other group
is same combination with bovine culture media (CR1aa). As a results, interspecies NT embryos
were not blocked at 2-cell stage in any type medium. A majority of the interspecies NT embryos
showed abnormal development in bovine culture media. In order to overcome abnormality and
blocking at 8-cell stage, we examine that antioxidant affected on developmental potential. There
are four different Experiments. First experiment was cultured in CR1aa with 5.5μM BME and
second experiment was cultured in CR1aa with 11μM BME. Each blastocysts rates were 2.2% and
3.8%, respectively. And, third experiment was cultured in CR1aa with 11μM BME plus 2mM GTH and
fourth experiment was cultured in CR1aa with 1μM BME plus 100μM GTH. Each blastocyst rates
were 1.1% and 3.6%, respectively. To know the mechanisms of Xist gene expression, we need to
examine the exact timing of its onset. Earlier studies reported that Xist is expressed during
development beginning from the 4-cell stage in mouse female preimplantation embryos. but we
could not find it at the 4-cell stage interspecies embryo. The interspecies embryo did not show
similarity of mouse embryo''s expression. β-actin mRNA was detected in all mouse embryo but FGF4, IGF2 receptor mRNA present in mouse 8-cell stage embryo. Expression of several gene, such
as FGF4, IGF2 receptor, was only detected in normal mouse embryo and poly(A) polymerase in
bovine gene was only detected in interspecies NT embryo. Thus, we think that the gene expression
in interspecies NT embryo was abnormal.
Language
English
URI
http://dcollection.snu.ac.kr:80/jsp/common/DcLoOrgPer.jsp?sItemId=000000053968

http://hdl.handle.net/10371/67614
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College of Agriculture and Life Sciences (농업생명과학대학)Dept. of Food and Animal Biotechnology (식품·동물생명공학부)Theses (Master's Degree_식품·동물생명공학부)
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