• 한국수정란이식학회:학술대회논문집
• /
• 한국수정란이식학회 2005년도 제5회 발생공학 국제심포지엄 및 학술대회
• /
• pp.113-113
• /
• 2005

• 한국수정란이식학회:학술대회논문집
• /
• 한국수정란이식학회 2006년도 The 6th Intermational Symposium on Developmental Biotechnology
• /
• pp.20-21
• /
• 2006

• 한국수정란이식학회:학술대회논문집
• /
• 한국수정란이식학회 2005년도 제5회 발생공학 국제심포지엄 및 학술대회
• /
• pp.146-146
• /
• 2005

• 한국수정란이식학회:학술대회논문집
• /
• 한국수정란이식학회 2003년도 제3회 발생공학 국제심포지움 및 학술대회
• /
• pp.111-111
• /
• 2003

• 한국수정란이식학회:학술대회논문집
• /
• 한국수정란이식학회 2006년도 The 6th Intermational Symposium on Developmental Biotechnology
• /
• pp.113-113
• /
• 2006

• 한국수정란이식학회:학술대회논문집
• /
• 한국수정란이식학회 2001년도 추계학술대회 및 정기총회
• /
• pp.35-35
• /
• 2001

• 한국수정란이식학회지
• /
• 제26권3호
• /
• pp.171-180
• /
• 2011

In this study I report that in vitro development rates of bovine nuclear transfer embryos activated either with boar sperm cytosolic factor (SCF) or with ionomycin followed by cycloheximide (CHX) and subsequent in vivo developmental rates after embryo transfer are related to blastocyst quality as evaluated by apoptosis analysis. SCF was extracted from porcine semen then purified for post-activation injection after nuclear transfer. The optimal timing for SCF injection was determined to be at least 22 h post-IVM for parthenogenetic activation of bovine oocyies. A total of 364 oocytes were successfully enucleated and 268 (73.6%) fused and were injected with SCF. The survival rate of fused and injected embryos was 109/113 (96.5%) after 2 h. The cleavage rates of nuclear transfer embryos after 3 d of culture in the ionomycin/CHX treated group were significantly higher than those of the SCF-activated group (93.3% vs 81.7%, p<0.01, respectively). However, at 7 d and 9 d there was no significant difference between the total developmental rates to blastocyst for either treatment group. Total blastocyst cell numbers were also not significantly different between the two activation treatments (ionomycin/CHX: 149.5${\pm}$7.7 vs. SCF: 139.3${\pm}$4.4 cells). In contrast, the apoptotic levels in the SCF blastocysts were higher than those produced after the chemical treatment (28.2${\pm}$5.1% vs. 8.8${\pm}$0.6%, respectively). A total of 18 expanded or hatching blastocysts was transferred to nine synchronized recipients in each activation group; 5/9 (55.5%) and 2/9 (22.2%) were pregnant at 40 d in the ionomycin/CHX treatment and SCF activated group, respectively. However, only one went to term in the ionomycin/CHX treatment while none of the pregnancies from the SCF group were maintained by 90 d. In conclusion, these results suggest that SCF derived from different species is a limited activator to be used for activation after bovine nuclear transfer in lieu of a chemical activation protocol.

• Reproductive and Developmental Biology
• /
• 제32권4호
• /
• pp.249-253
• /
• 2008

Interspecies somatic cell nuclear transfer (iSCNT) is a valuable tool for studying the interactions between an oocyte and somatic nucleus. The object of this study was to investigate the developmental competence of in vitro-matured porcine oocytes after transfer of the somatic cell nuclei of 2 different species (goat and rabbit). Porcine cumulus oocytes were obtained from the follicles of ovaries and matured in TCM-199. The reconstructed embryos were electrically fused with 2 DC pulses of 1.1kV/cm for $30{\mu}s$ 0.3M mannitol medium. The activated cloned embryos were cultured in porcine zygote medium-3 (PZM-3), mSOF or RDH medium for 7 days. The blastocyst formation rate of the embryos reconstructed from goat or rabbit fetal fibroblasts was significantly lower than that of the embryos reconstructed from porcine fetal fibroblast cells. However, a significantly higher number of embryos reconstructed from goat or rabbit fetal fibroblasts cultured in mSOF or RDH, respectively, developed to the morular stage than those cultured in PZM-3. These results suggest that goat and bovine fetal fibroblasts were less efficacious than porcine-porcine cloned embryos and that culture condition could be an important factor in iSCNT. The lower developmental potential of goat-porcine and porcine-bovine cloned embryos may be due to incompatibility between the porcine oocyte cytoplasm and goat and bovine somatic nuclei.

• Reproductive and Developmental Biology
• /
• 제31권2호
• /
• pp.109-113
• /
• 2007

This study was conducted to evaluate the effect of cytochalasin B (CB) treatment in the activation medium on the development of somatic cell nuclear transfer (SCNT) rat embryos. Fetal fibroblast cells were isolated from a Day 14.5 fetus, and the oocytes for recipient cytoplasm were recovered from 4-week old Sprague Dawley rats. After enucleation and nuclear injection, the reconstructed oocytes were immediately exposed to activation medium consisting of 10 mM $SrCl_2$ with or without CB for 4 hr, and formation of pseudo-pronucleus (PPN) was checked at 18 hr after activation. Then, they were transferred into day 1 pseudopregnant recipients (Hooded Wistar) or cultured for 5 days to check their developmental competence in vivo or in vitro. The number of PPN was not affected by CB treatment during the activation. However, CB treatment supported pre-implantation development of rat SCNT embryos. Embryos generated by the procedures of SCNT were also capable of implanting, with 1 implantation scar found from a recipient following the transfer of 87 SCNT embryos to four foster mothers. The result of the present study shows that rat SCNT embryo can develop to post-implantation stage following treatment with CB.

• 한국수정란이식학회지
• /
• 제11권2호
• /
• pp.95-101
• /
• 1996

The present study investigated the effect of treatment with nocodazole on the efficiency of cell cycle synchronization and development of mouse 4-cefl embryos. In addition, developmental ability of reconstituted embryos that received nuclei from 4-cell embryos synchronized with nocodazole at M phase was examined in vitro and in vivo. (1) When 4-cell blastorneres exposed to culture medium containing l$\mu$g /ml nocodazole for 2, 4, 6 and 8 hrs, 40% (40/10l), 75% (l19/159), 85% (87/102) and 97% (155/160) of nuclei were synchronized at M phase, respectively. (2) Treated with nocodazole for 4 hrs, the proportion of 4-cell embryos developed to blastocysts (98%, 60/61) was not significanUy different from that of the control embryos (98%, 196/201). However, the developmental ability of 4-cell embryos treated for 8 (87%, P<0.05)and 12 hrs (76%, P