Development of an interspecies somatic cell nuclear transfer system for establishing innovative medical treatment
새로운 치료기술 개발을 위한 이종간 체세포 핵이식 기술 확립

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서울대학교 대학원
인간 태줄 상피 세포Human cord cell소 난자Bovine oocyte이종간 체세포 핵이식Intergenetic somatic cell nuclear transfer배아간 줄기 세포Blastocyst formation배반포 발육Cytogenetic analysis염색체 검사Mitochondrial DNA
Thesis (master`s)--서울대학교 대학원 :농생명공학부,2003.
Recently, cell and tissue therapies with employing somatic cell nuclear
transfer (SCNT) have been suggested to develop innovative medical technology,
which greatly contributes to overcome degenerative diseases.
After the successful birth of cloned sheep, ¡°Dolly¡±, SCNT is of
significantly greater commercial and research importance. However, various
concerns on human SCNT, which might cause human cloning, have been made.
The objective of my master thesis was to develop an efficient SCNT, which
could develop an alternative therapeutic cloning technology without sacrificing
human oocytes. To achieve this purpose, I attempted to establish an interspecies
nuclear transfer of human somatic cell into enucleated bovine oocytes.
In the first series of experiment, I attempted to select the most efficient
somatic cell type and preparation, fusion medium and culture strategy for
interspecies clone embryos.
As results, SCNT of fetal cord fibroblasts yielded more (P=0.0006) 2-cell
embryos than SCNT of adult fibroblasts. A significant (P<0.05) batch effect of
cord fibroblast on preimplantation development after SCNT was found and serum
starvation greatly improved preimplantation development, regardless of the
batches. No significant effect was observed after change of fusion medium (Ca2+-
free mannitol solution) to Ca2+-containing mannitol or sorbitol solution, while
embryo culture system greatly influenced the development. A continuous culture
in modified synthetic oviduct fluid medium used in bovine embryo culture only
supported blastocyst formation of interspecies clone embryos.
In the second series of experiment, I further optimized the interspecies SCNT
developed in my previous experiment by changing cord fibroblast cell batches and
reconstruction method. Total 1,742 bovine oocytes were provided for SCNT.
Reconstruction of a fibroblast by single DC pulse of 1.9 to 2.1 kV/cm for 20
¥ìsecond yielded better rate of fusion (30 to 56%) and, development to the 2-
cell (27 to 36%), 8-cell (7 to 14%), 16-cell (5 to 7%) and morula to blastocyst
(3 to 5%) stages. Results from karyotyping and mitochondrial DNA analysis
demonstrated that 56% of karyotyped embryos had presumptively normal human
chromosome complements consisting of 23 pairs of autosomes with sex chromosome,
while only bovine mtDNA was detected.
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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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