The development of embryos reconstructed by nuclear transfer is dependent upon numerous factors including the type of recipient cell, method of enucleation, the type of donor cell, method of reconstruction, activation, the cell cycle stage of both the donor nucleus and the recipient cytoplasm and the method of culture of the reconstructed embryos. Many of these points which have been reviewed extensively elsewhere (Sun and Moor, 1995; Colman, 1999; Oback and Wells, 2002; Renard et al., 2002; Galli et al., 2003b), here we will concentrate on main area, the production of suitable cytoplast and nuclear donor, nuclear-cytoplasmic coordination, oocyte activation, culture of reconstructed embryos, and the effects that this may have on development.

This work was undertaken in order to study the developmental competence of nuclear transfer cat embryo with fetal fibroblast and adult skin fibroblast as donor nuclei. Oocytes wererecovered by mincing the ovaries in Hepes-buffered TCM199 and selected the cumulus oocyte complexes (COCs) with compact cumulus cell mass and dark. Homogenous ooplasm were cultured for maturation in TCM199 + 10% fetal bovine serum (FBS) for 12 hours and used as a source of recipient cytoplast for exogenous somatic nuclei. In Experiment 1, we evaluated the effect donor cell types on the reconstruction and development of cloned embryos. Fusion, first cleavage and blastocyst developmental rate was not different between fetal fibroblast and adult skin cell (71.2 vs. 66.8; 71.0 vs. 57.6; 4.0 vs. 6.1 %, P<0.05). In Experiment 2, cloned embryos were surgically transferred into the oviducts of recipient queens. One of seven recipient queens was delivered naturally 2healthy cloned cats and 1 stillborn from fetal fibroblast cell of male origin after 65 days embryo transfer. One of three recipient queens was delivered naturally 1 healthy cloned cat from adult skin cell of female after 65 days embryo transfer. The cloned cats showed genotypes identical to the donor cell lines, indicating that adult somatic cells can be used for feline cloning.

본 연구는 토천궁의 종묘비 절감과 증식율을 향상시키기 위한 삽목번식법을 이용하고자 할 때 삽수의 발근과 생육에 미치는 요인을 알아보고자 실험하였으며 얻어진 결과는 아래와 같다. 1. 발근율은 Perlite(87.5%) 〉 Perlite 2+ Vermiculite 1 (63.3%) 〉 Sand(33.8%) 〉 Upland soil(14.4%) 순이었으며, 생장은 통기성이 좋아 배지 내에 수분과 공기의 출입이 원활한 Perlite 단용처리에서 가장 좋았다. 2. 관수방법과 관수량 실험에서 저면관수로 30분 간격을 두고 15분 관수를 하는 것이 생육이 가장 좋았다. 3. 줄기 삽수시 생육 초기에는 물만을 공급하고 20일경부터 원예연구소 표준액의 농도를 1/2배로 하여 처리하는 것이 발근과 생육에 더 유리한 것으로 나타났다. 4. 줄기 삽수시 발근 및 생장에 적합한 온도는 30℃=25℃ > 20℃ 순이었고, 일장은 영향을 주지 않았다.