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한정 배양액에서 포유동물 수정란의 체외 발생에 인, 아미노산 및 BSA가 미치는 영향
김종흥,김병기 한국생명과학회 1996 생명과학회지 Vol.6 No.3
본 연구는 포유동물 수정란의 체외발생에 인, 아미노산과 BSA가 미치는 영향을 조사하기 위하여 체외성숙 및 수정된 소의 수정란을 한정 무단백 배양액에서 배양하였다. 배양액에 포함된 인의 농도를 0.00, 0.10. 0.35, 1.05와 2.10mM로 조정하였을 때 0.00부터 1.05mM 농도에서는 수정후 48시간 까지 수정란 발생에 영향을 미치지 않았고, 수정후 96시간과 114에서의 8세포기와 상실배 발생이 0.35mM에서 유의적(P<0.05)으로 붕가하였다. 19종 아미노산의 첨가는 수정후 96, 144, 192시간에 각각8세포기(49-50%), 상실배(38-40%), 배반포(29-32%)발생을 유의적(P<0.05)으로 증가하였으나, glutamine단독 첨가는 영향이 없었다. BSA첨가는 첨가시간에 관계없이 수정후 48, 96, 144, 192시간에서 각각 2세포기, 8세포기, 상실배와 배반포의 발생을 증가시켰다. The aim of this study was to evaluate the effects of phosphate, aimno acid, and BSA on in vitro development of mammalian embryos. In vitro-matured and -fertilized(IVM/IVF) bovine embryos were cultured in simple, chemically defined, protein-free medium(mTLP-PVA0. When the phosphate concentration of mTLP-PVA supplemented with 19 amino acid were adjusted to 0.0, 0.10, 0.35, 1.05 and 2.10mM by the concentration of sodium phoshpate, there were no significant different in development ability of IVM/IVF bovine embryos cultured in the medium containing from 0.00 to 1.05mM phosphate until 48 hours post-insemination, However, proportion of embryos developing to $$8-cell and morula at 96 and 144 hours post- insemination, respectively, was significantly increased in the medium with o.35 mM phosphate(p<0.05). There was significant difference between O.10(18%)-0.35(24%)mM phosphate and 1.05(13%)-2.10(1%)mM phosphate in supporting development to blastocyst(p<0.05). When IVM/IVF bovine embryos were cultured in the medium supplemented with 19 amino acids, significant different was observed in the proporton of embryos reaching $$8-cell(49-50%), morula(38-40%) and blastocyst (29-32%) stages at 96, 144, and 192 hours post-insemination, respectively(p<0.05). Glutamine alone had no benefit on embryo development. When BSA was added to mTLP-PVA with 0.35mM phosphate, glutamine and 19 amino acids at 8, 48, 120 hours post-insemination, BSA significantly enhanced the development ability ofb embryos reaching $$2-cell (74-77%), $$8-cell (49-53%), morula(43-47%), and blastocyst(38-42%) stages at 48, 96, 144, and 192 hours post-insemination, respectively, regardless of the time of BSA addition.
Kim, J. H.,Lee, C. S.,Kim, B. K. 동의대학교 기초과학연구소 1997 基礎科學硏究論文集 Vol.7 No.1
본 연구는 배양액의 삼투압과 비타민이 체외 배발생에 미치는 영향을 조사하기 위하여 소의 미성숙 난포란을 체외성숙 및 수정 후 무단백 한정배양액에서 체외배양하였다. 0.35mM phosphate와 19종 아미노산이 포함된 무단백 한정배양액의 삼투압을 NaCl농도에 의하여 달리하였을 때 소 수정란의 발생능력은 265mOsm와 290mOsm에서 유의적으로 증가하였다 (p$lt;0.05). 수정 후 120시간에 5.56mM포도당이 포함된 새로운 배양액에서 배발생을 계속시켰을 때 상실배는 290mOsm과 315mOsm에서 유의적으로 증가하였으며, 배반포는 290mOsm배양액에서 유의적으로 증가하였다. Glutamine과 아미노산이 첨가된 배양액에서 비타민은 소수정란의 발생에 영향을 미치지 않았다.
Kim, Jong-Heung,Kim, Byung-Ki 동의대학교 기초과학연구소 1997 基礎科學硏究論文集 Vol.7 No.1
The aim of this study was to evaluate the effects of phosphate, amino acid, and BSA on in vitro development of mammalian embryos. In vitro-matured and -fertilized(IVM/IVF) bovine embryos were cultured in simple, chemically defined, protein-free medium(mTLP-PVA). When the phosphate concentration of mTLP-PVA supplemented with 19 amino acid were adjusted 0.0, 0.10, 0.35, 1.05 and 2.10mM by the concentration of sodium phoshpate, there was no significant different in development ability of IVM/IVF bovine embryos cultured in the medium containing from 0.00 to 1.05mM phosphate until 48 hours post-insemination. However, proportion of embryos developing to ≥8-cell and morula at 96 and 144 hours post-insemination, respectively, was significantly increased in the medium with 0.35 mM phosphate(p$lt;0.05). There was significant difference between 0.10(18%)-0.35(24%)mM phosphate and 1.05(13%)-2.10(1%) mM phosphate in supporting development to blastocyst(p$lt;0.05). When IVM/IVF bovine embryos were cultured in the medium supplemented with 19 amino acids, significant different was observed in the proportion of embryos reaching ≥8-cell(49-50%), morula(38-40%) and blastocyst (29-32%) stages at 96, 144, and 192 hours post-insemination, respectively(p$lt;0.05). Gultamine alone had no benefit on embryo development. When BSA was added to mTLP-PVA with 0.35mM phosphate, glutamine and 19 amino acids at 8, 48, 120 hours post-insemination, BSA significantly enhanced the developmental ability of embryos reaching ≥2-cell (74-77%), ≥8-cell (49-53%), morula(43-47%), and blastocyst(38-42%) stages at 48, 96, 144, and 192 hours post-insemination, respectively, regardless of the time of BSA addition