This study was designed to investigate the effect of essential amino acids (EAA) and/or non-essential amino acids (NEAA) on the development of parthenogenetic and somatic cell nuclear transfer (SCNT) porcine embryos in vitro. To evaluate the timing of amino acids supplementation, activated oocytes w...
This study was designed to investigate the effect of essential amino acids (EAA) and/or non-essential amino acids (NEAA) on the development of parthenogenetic and somatic cell nuclear transfer (SCNT) porcine embryos in vitro. To evaluate the timing of amino acids supplementation, activated oocytes were cultured in NCSU23-PVA with EAA, NEAA or NEAA+EAA (AAs) during specific periods as below: EAA, NEAA or AAs were supplemented during Day 0 to 6 (whole culture period: ALL), Day 2 to Day 6 (post-maternal embryonic transition period: POST-MET), Day 5 to Day 6 (post-compaction period: POST-CMP), Day 0 to Day 2 (pre-maternal embryonic transition period: PRE-MET), or Day 0 to Day 4 (post-compaction period: PRE-CMP). Supplementation of NEAA decreased cleavage rates in PRE-MET and PRE-CMP and also decreased blastocyst rates in POST-CMP. On the other hand, EAA significantly enhanced blastocyst formation rate in POST-MET and no detrimental effect on embryonic development in other groups. Interestingly, NEAA and EAA had synergistic effect when they were supplemented to the medium during whole culture period. Supplementation of AAs also enhanced SCNT porcine embryo development whereas BSA-free medium without AAs could not supported blastocyst formation of SCNT embryos. In conclusion, existence of EAA and NEAA in defined culture medium variously influences the development of parthenogenetic and SCNT porcine embryos, and their positive effect are only occurred when both EAA and NEAA are supplemented to the medium during whole culture period. Additionally, AAs supplementation enhances the blastocyst formation of SCNT porcine embryos when they are cultured in the defined condition.
This study was designed to investigate the effect of essential amino acids (EAA) and/or non-essential amino acids (NEAA) on the development of parthenogenetic and somatic cell nuclear transfer (SCNT) porcine embryos in vitro. To evaluate the timing of amino acids supplementation, activated oocytes were cultured in NCSU23-PVA with EAA, NEAA or NEAA+EAA (AAs) during specific periods as below: EAA, NEAA or AAs were supplemented during Day 0 to 6 (whole culture period: ALL), Day 2 to Day 6 (post-maternal embryonic transition period: POST-MET), Day 5 to Day 6 (post-compaction period: POST-CMP), Day 0 to Day 2 (pre-maternal embryonic transition period: PRE-MET), or Day 0 to Day 4 (post-compaction period: PRE-CMP). Supplementation of NEAA decreased cleavage rates in PRE-MET and PRE-CMP and also decreased blastocyst rates in POST-CMP. On the other hand, EAA significantly enhanced blastocyst formation rate in POST-MET and no detrimental effect on embryonic development in other groups. Interestingly, NEAA and EAA had synergistic effect when they were supplemented to the medium during whole culture period. Supplementation of AAs also enhanced SCNT porcine embryo development whereas BSA-free medium without AAs could not supported blastocyst formation of SCNT embryos. In conclusion, existence of EAA and NEAA in defined culture medium variously influences the development of parthenogenetic and SCNT porcine embryos, and their positive effect are only occurred when both EAA and NEAA are supplemented to the medium during whole culture period. Additionally, AAs supplementation enhances the blastocyst formation of SCNT porcine embryos when they are cultured in the defined condition.
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제안 방법
The activated oocytes were treated for 5~6 h in North Carolina State University-23 (NCSU-23) medium supplemented with 5 //g/ml cytochalasin B and 1 mg/ml PVA. The oocytes were then washed nine times with culture medium and cultured in 20-“1 drops (10 ~15 oocytes per drop) of the appropriate culture medium for 7 days at 38.5 ℃ and 5% CO, To evaluate the timing of amino acids supplementation, activated oocytes were cultured in NCSU23- PVA with EAA, NEAA or NEAA+EAA (AAs) during specific culture periods as below: EAA, NEAA or AAs were supplemented during Day 0 to 6 (whole culture period group: ALL), Day 2 to Day 6 (post-maternal embryonic transition period group: POST-MET), Day 5 to Day 6 (post-compaction period group: POST-CMP), Day 0 to Day 2 (pre-matemal embryonic transition period group: PRE-MET), or Day 0 to Day 4 (pre-compac- tion period group: PRE-CMP). In each group, 150—160 oocytes were evaluated.
, 2000). To evaluate the timing of amino acids supplementation, porcine parthenotes were cultured in EAA, NEAA or EAA+NEAA (AAs) supplemented medium during specific culture periods (whole culture period, post- maternal embryonic transition period, post-compaction period, pre-matemal embryonic transition period, or post-compaction period). In this study, we investigated the effects of amino acid components on the development of parthenogenetic and SCNT porcine embryos to the blastocyst stage to optimize defined culture system for the production of porcine embryos in vitro.
데이터처리
Experiments were repeated 3—5 times, and mean values were analyzed by ANOVA analysis of variance. Difference at p<0.
성능/효과
In conclusion, existence of EAA and NEAA in defined culture medium variously influenced in vitro development of par- thenogenetic and SCNT porcine embryos, and their positive effect were only occurred when both EAA and NEAA were supplemented to the medium during whole culture period. In addition, AAs supplementation was essential for the development of SCNT porcine embryos when they were developed in defined culture condition.
The activation process for reconstructed embryos was the same as that used for parthenogenetic embryos. The reconstructed oocytes were washed with culture medium nine times and cultured in 20 *1 drops (10~15 oocytes per drop) of NCSU-23 with either 1 mg/ml PVA (with or without AAs) or 4 mg/ml BSA for 7 days at 38, 5 3C in 5% CO2 in air and the number of total and hatching blastocysts were counted (Fig. 2), In total, 804 SCNT embryos were evaluated (250—300 embryos per group).
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