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전기자극법에 의한 생쥐 2세포기란의 분할구 융합에 관한 연구
양부근,한상익,김정익,Yang, B.Y.,Han, S.I.,Kim, C.I. 대한생식의학회 1991 Clinical and Experimental Reproductive Medicine Vol.18 No.1
To find out the suitable method for blastomeres fusion of mouse 2-cell embryo using electric stimuli, these studies were carried out with various voltages (1.0 KV, 1.2 KV, 1.5 KV, 1.7 KV and 2.0KV), pulse duration times($50{\mu}\;sec$, $75/{\mu}\;sec$, $100{\mu}\;sec$) and different fusion solutions. In addition, the fused embryos were cultured for 72-80hr to observe their subsequent development. These results were summarized as follows: 1. The proportion of the fused embryos were 50.8%(34/67), 60.7%(34/56), 70.6%(48/68), 66.7% (48/72) and 85.3% (58/68) after stimuli of 1.0KV, 1.2KV, 1.5KV, 1.7KV and 2.0KV for $100{\mu}\;sec$ with 2 times, and the electric stimulation at 2.0KV(85.3%) was the most effective voltage on the blastomere fusion. 2. For in vitro development, blastocysts of the fused embryos were cultured for 72-80hrs in $M_{16}$ medium. The group(52.1%) treated with 1.5KV for $100{\mu}\;sec$ with 2 times showd higher development rates than those any other group. However, these results were not corresponded to those of the rates of blastomere fusion. 3. There were no significant differences among the rates of blastomeres fusion to 50(70.6%), 75(71.9%), and 100(78.0%) ${\mu}sec$ stimulation at 1.5KV with two times. However, the development rates of the fused embryo in vitro were 52.1%(25/48), 28.3%(13/46) and 9.4%(3/32) at the above conditions, and the development rates of fused embryo increased as the pulse duration times increased. 4. The rates of the blastomeres fusion were 38.9% (28/72) or 70.6% (48/68) in electrolyte (PBS) or non-electrolyte(0.3M mannitol) solution. The development rates of the fused embryo were 32.1% (9/28) or 52.1%(25/48) in the above fusion solutions, and non-electrolyte-treated group showed higher development rates of embryo than that of electrolyte-treated group.
Catalase와 $\beta$-Mercaptoethanol이 돼지 태아섬유아세포 Clonal Lines의 배양에 미치는 영향
권대진,박선영,박춘근,양부근,김정익,정희태,Kwon D. J.,Park S. Y.,Park C. K.,Yang B. K.,Kim C. I.,Cheong H. T. 한국수정란이식학회 2004 한국동물생명공학회지 Vol.19 No.3
본 연구에서는 clonal cell lines을 효율적으로 확립할 수 있는 방법을 제시하기 위하여 배양액 내에 catalase와 $\beta$ME 첨가가 clonal cell line 확립 효율에 미치는 영향을 검토하였다. 임신 50일령의 암퇘지에서 얻은 태아섬유아세포를 2회 passage한 후 동결 보관하였다가 실험에 이용하였다. 단일세포를 catalase나 $\beta$ME가 첨가된 배양액이 들어 있는 96-well dish로 옮겨 배양하였다. 단층이 형성된 세포는 4-well dish로 옮겨주어 배양하였으며, 이후 2회 이상 passage가 가능한 세포를 clonal line이 확립된 세포로 판정하였다. 실험 1에서 clonal line 확립효율에 미치는 catalase와 $\beta$ME의 효과를 검토하기 위하여 세포를 100ng/$m\ell$) catalase와 100nM $\beta$ME가 첨가된 DMEM액 내에서 배양하여 clonal line 확립효율을 검토하였다. $\beta$ME 첨가 시 clonal line 확립효율이 8.3%로 대조구의 3.2%에 비해 유의적으로 높게 나타났다.(P<.0.05). 그러나 catalase 첨가 시(3.6%)에는 대조구와 차이가 없었다. 실험 2에서 catalase 농도(0, 10, 100, 1,000ng/$m\ell$)에 따른 clonal line 확립효율을 검토하였다. 대조구에 비하여 모든 처리구에서 clonal line 확립 효율에 대한 첨가효과가 없었다(0-2.6%). 실험 3에서 $\beta$ME 농도(0, 10, 100, 1,000 nM)에 따른 clonal line 확립효율을 검토하였다. 100 nM의 $\beta$ME 첨가 시 clonal line 확립 효율이 9.4%로 대조구 및 타 처리구(0-l.6%)보다 유의적으로 높게 나타났다(P<0.05). 본 연구 결과 배양액 내 catalase 첨가는 확립효율에 영향을 미치지 않지만, 100nM의 $\beta$ME 첨가로 clonal cell line의 확립효율을 향상시킬 수 있는 것으로 나타났다. This study was performed to examine the effects of catalase and $\beta$-mercaptoethanol ($\beta$ME) on the establishment of clonal lines from porcine fetal fibroblast cells. Fibroblasts derived from a pig fetus (Day 50) were passaged two times before use. A single cell was seeded in 96-well plates and cultured in medium supplemented with or without catalase or $\beta$ ME. Cell colonies were passaged two times into 4-well dish. Cell lines with proliferating potential were classified as an established clonal cell line. In experience 1, the establishment efficiencies were examined by addition of catalase (100ng/$m\ell$) or $\beta$ME (100 uM) in culture medium. The establishment efficiency of $\beta$ME-added group (8.3%) was significantly higher than that of control group (3.2%, P<0.05). However, catalase did not have a positive efffct on the establishment efficiency. In experience 2, the establishment efficiencies were examined by addition of different concentrations of catalase (0-1,000 ng/$m\ell$) in culture medium. However, establishment efficiencies were not different among the different concentrations of catalase (0-2.6%). In experience 3. the establishment efficiencies were examined by addition of different concentrations of $\beta$ME(0-1,000 uM) in culture medium. The establishment efficiency was significantly higher in 100 uM $\beta$ME-added group (9.4%) compare to others (0-1.6%). The result of present study shows that the establishment efficiency of clonal cell lines can be enhanced by the culture in media supplemented with 100uM $\beta$ME. However, catalase did not have a positive effect on the establishment efficiency.
한우도체의 부분육 수율에 영향을 미치는 성 , 도체중 및 도체평가 형질들의 효과
이재윤,김종복,신종서,양부근,홍병주 ( J . Y . Lee,J . B . Kim,J . S . Shin,B . K . Yang,B . J . Hong ) 한국축산학회 1997 한국축산학회지 Vol.39 No.2
The data of 550 beef tattles collected after grading in Seoul Whole Sale Market, NLCF from September, 1995 to March, 1996 were analyzed to estimate effects of sex, carcass weight and some carcass traits on retailed cut percentage. Correlation between lean and supreme meat percentages was 0.741 and correlation between lean and top grade meat percentages was 0.939. For steers, lean meat, bone, supreme meat, and top grade meat percentages were 65.31, 12.80, 15.15, and 37.74%, respectively, which were lower than those of bull which were 67.27, 13.09, 16.09 and 12.41%, respectively. However, fat percentage of 21.88% and rib percentage of 12.41% were higher than those of bulls which were 19.41 and 12.06%, respectively. In cow, bone percentage of 12.30% was low but ratio of muscle to bone of 1.84 was high. The grade 1 of meat quality obtained higher fat yield and ratio of fat to bone than the grade 2 and 3 of meat quality. The grade A of meat yield obtained higher lean meat percentage, top grade meat percentage, ratio of muscle to bone, and ratio of muscle to fat which were respectively 67.03, 39.09, 5.26 and 3.56% than the grade B and C of meat yield.
박춘근,정희태,김종흥,이상찬,양부근,김정익,Park, C.K.,Cheong, H.T.,Kim, J.H.,Lee, S.C.,Yang, B.K.,Kim, C.I. The Korean Society for Reproductive Medicine 1998 Clinical and Experimental Reproductive Medicine Vol.25 No.3
The objective of this study was to test the effect of catalase on penetration in vitro by spermatozoa preincubated with xanthine and/or xanthine oxidase. The penetration rates were, significantly (p<0.05) higher in spermatozoa preincubated without (66 and 38%) than with (40 and 15%) catalase for 0 and 30 min. When spermatozoa were preincubated and inseminated in medium with xanthine, the penetration rates were significantly higher (p<0.05) in medium with (68, 70 and 49% for 0, 30 and 60 min) than without (33, 41 and 19% for 0, 30 and 60 min) catalase. However, in oocytes were' inseminated with spermatozoa pre incubated with or without catalase in the presence of xanthine oxidase, no decrease in penetrations rates were observed for up to 60 min of preincubation. In another experiment, the penetration rates were significantly (p<0.00l) higher in medium with (75, 55 and 52%) than without (14, 4 and 8%) catalase when oocytes were inseminated with spermatozoa preincubated for 0, 30 and 60 min in the presence of xanthine plus xanthine oxidase. On the other hand, The rate of polyspermy in oocytes penetrated in medium without catalase in the presence of xanthine or xanthine plus xanthine oxidase decreased with time of spermatozoa preincubation. However, no differences were observed in polyspermy rates in the medium with xanthine oxidase alone despite presence of catalase. These results indicate the advantages of spermatozoa pre incubated with xanthine plus xanthine oxidase in the presence of catalase to increase penetration potential and with suppressed polyspermy in porcine. 본 연구는 xanthine과 xanthine oxidase가 첨가된 배양액내에서 전배양된 돼지동결정액의 수정능력에 있어서 catalase의 영향을 검토하였다. 체외수정을 위한 기본 배양액내에서 0 또는 30분간 전배양한 정자는 catalase를 첨가 (40 및 15%)한 경우 보다는 무첨가 (66 및 38%)시 유의적으로 높은 정자침입율을 나타냈지만 (p<0.05), 배양액내에 xanthine을 첨가해 정자를 0, 30 및 60분간 전배양했을 때에는 catalase 무첨가 (33, 41 및 19%) 보다는 첨가시 (68, 70 및 49%) 유의적으로 높은 정자침입율이 인정되었다 (p<0.05). 그러나 xanthine oxidase를 첨가하여 정자의 전배양을 행하지 않은 경우는 catalase의 첨가 (13%) 보다는 무첨가 (51%)시 유의적으로 높은 정자침입율을 나타냈지만 (p<0.01), 전배양(30 및 60분)후에는 catalase의 존재유무와 정자전배양시간에 관계없이 매우 낮은 정자침입율 $(10\sim21%)$을 나타냈다. xanthine과 xanthine oxidase를 동시에 첨가하여 0, 30 및 60분간 정자를 전배양한 경우 catalase의 무첨가 (14, 4 및 8%)보다 첨가 (75, 55 및 52%)시 유의적으로 높은 정자침입율을 나타냈다 (p<0.001). 한편, 다정자침입율은 xanthine, xanthine+xanthine oxidase의 첨가시 정자전배양기간이 길어짐에 따라 감소하였으며, catalase의 첨가보다는 무첨가시 낮은 다정자침입율을 나타냈다. 본 연구의 결과로부터 xanthine과 xanthine oxidase를 동시에 첨가시 catalase의 존재는 정자의 전배양후에도 다정자침입을 억제하면서 수정능력의 유지를 위해 매우 효과적인 것으로 추측되었다.
정희태(H . T . Cheong),임석기(S . K . Im),이준희(J . H . Lee),박춘근(C . K . Park),양부근(B . K . Yang),김정익(C . I . Kim) 한국축산학회 1996 한국축산학회지 Vol.38 No.6
This study was conducted to examine the possibility of cell cycle synchronized nuclear transfer and the effect of the early cell cycle stage of donor embryo on the development of nuclear transplant bovine embryos. Donor embryos were treated with nocodazole, then monitored their nuclear phase and cleavage after release from the nocodazole treatment. Blastomeres cleaved within 3 hr and noncleaved ones were transferred into enucleated oocytes. Some nuclear transplant embryos were fixed to investigate their chromatin structures. Comparatively high proportions(56.0-58.6%) of blastomeres arrested at mitotic phase(M-phase) were obtained following treatments with 0.5∼1.0㎍/㎖ of nocodazole for 16 hr. Cleavage rates of blastomeres within 3 hr after release from nocodazole treatment were average 64%(188/294). After nuclear transfer, 13%(11/85) of embryos were developed to blastocysts following transplantation of blastomeres within 1.5 hr post cleavage(hpc), but 7%(5/74) and 5%(3/63) were developed to blastocysts following transplantation of blastomeres at 3 hpc and noncleaved ones. About 40% of 1.5 hpc nuclear transplant embryos have a single chromatin clump, whereas only 25.0% of 3.0 hpc nuclear transplant eanbryos have a single chromatin clump and the rest have 2 or multi chromatin clumps. From the result of this study, it was suggested that the developmental potential of nuclear transfer bovine embryos may be enhanced with transfer of early cell cycle stage blastomeres immediately after cleavage. It was also suggested that the cell cycle synchronization of bovine embryonic cells at GI phase is very difficult.
유전 및 육종 : 분만년도, 계절 및 산차가 한우의 비유곡선에 미치는 영향
황정미 ( J. M. Hwang ),최재관 ( J. K. Choi ),전기준 ( K. J. Jeon ),나기준 ( K. J. Na ),여인서 ( I. S. Yuh ),양부근 ( B. K. Yang ),이채영 ( C. Y. Lee ),김종복 ( J. B. Kim ) 한국축산학회 2002 한국축산학회지 Vol.44 No.6
This study was conducted to investigate the effect of environmental factor on milk yield and to estimate lactation curve in Korean cattle. The data for milk yields were collected from 118 cows from 1997 to 2000 at National Livestock Research Institute in Daekwanryoung, Kangwon-do. Average daily milk yields for 1st, 2nd, 3rd and 4th month after calving were 3.74㎏, 3.64㎏, 3.26㎏ and 2.99㎏. Average daily milk yield for the four months was 3.52㎏. The milk yields for cows calved in spring were larger than those calved in fall. Lactation curve of Korean cattle was y_n=2.4845n^0.1734e^-0.0060n. Peak milk yield was 3.75㎏ on 29.03 day after calving. The peak milk yields for multi-parous cows were larger than those of primiparous cows. The peak milk yields for multi-parous cows reached later than those for primiparous cows. The cows calved in spring had higher and earlier peak milk yields than those calved in fall had.