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성장생물학 : 지방과 근육 세포주의 단독 및 공동배양을 통한 세포형태학 및 세포물질 비교 연구
연성흠 ( Seong Heum Yeon ),조원모 ( Won Mo Cho ),백경훈 ( Kyung Hoon Baek ),송만강 ( Man Kang Song ),최창원 ( Chang Weon Choi ),황보순 ( Bo Soon Hwang ),박성권 ( Sung Kwon Park ) 한국동물자원과학회 ( 구 한국축산학회 ) 2012 한국축산학회지 Vol.54 No.2
본 연구는 기존 단독배양 위주로 이루어져온 세포배양 연구의 방법학적 한계의 극복과 대안을 제시하고자 지방과 근육세포주의 단독 및 공동배양에서 배양기법에 따른 지방 및 근육세포의 분화에 미치는 영향을 비교 조사하고자 실시하였다. 3T3-L1(지방세포) 및 L6(근육세포) 세포주는 성장배지인 10% FBS/DMEM(1% Pen- Strep solution 및 0.1% Fungizone 첨가) 하에서 48h 동안 단독배양 후 5% FBS/DMEM에서 배양하였다. 분화를 위한 단독 및 공동배양에서는 지방 및 근육세포 모두 분화유도물질 없이 2% FBS/DMEM으로 배양하였고, 공동배양에서는 0.4㎛ insert membrane을 사용하여 6 well plate 하단에 L6 cell을, 상단에는 3T3-L1 cell을 공생시켰다. 지방 및 근육세포 분화정도 측정은 세포별 형태학적 측정과 glycerol-3-phosphate dehydrogenase (GPDH) 및 creatine kinase(CK) 분석을 통해 조사되었다. 형태학적으로 볼때 3T3-L1 세포주는 공동배양보다 단독배양 시 분화가 더욱 잘 일어났고 L6 세포주의 경우 역으로 같았다. 세포물질분석에서는 분화배지 처리일(day 0)과 비교해 단독 및 공동배양 모두 지방세포 내 GPDH의 활성도가 유의적으로(P<0.05) 증가했음을 확인할 수 있었고 단독배양이 공동배양보다 유의적으로(P<0.05) 높은 수준의 GPDH 활성도를 보였다. L6 역시 마찬가지로 분화배지 처리일에 비하여 단독 및 공동배양 모두 CK 활성도가 유의적으로(P<0.05) 높았고, CK 활성도가 공동배양에서 유의적으로(P<0.05) 높게 나타남을 확인할 수 있었다. 이러한 결과는 기존 연구에서 이용된 단독 배양을 통한 세포 분화 결과 등은 생체와 비교 시 방법학적 한계로 인해 실제 생체 내에서는 그 분화정도가 매우 다를 것으로 생각되며, 이것은 앞으로 정확한 세포배양 결과 확보를 위해서는 단독배양보다는 공동배양기법을 사용해야 함을 의미한다. 향후 다양한 조건과 분화조절 물질들의 첨가를 통한 추가적인 공동배양실험이나 지방분화관련 분자생물학적 물질분석 등 다양한 실험 수행 시 보다 현실적이고 대량의 기초자료 확보가 가능할 것으로 판단된다. Present study was performed to investigate the effect of single and co-culture of adipocyte and muscle cell lines on cell differentiation. 3T3-L1(adipocyte) and L6(muscle) cell lines were single-cultured on the condition of 10% fetal bovine serum (FBS)/Dulbeco`s modified eagle`s medium(DMEM) for 48 h followed by culture within 5% FBS/DMEM as a growth media. Then, the growth media was replaced by differentiation media composed of 2% FBS/DMEM without additives in single- or co-culture of the 3T3-L1 and the L6 cells to induce differentiation of both cell types. In co-culture system, the 3T3-L1 and the L6 cells were grown in separated places by being seeded on a 0.4μm insert membrane and on the bottom of 6 well plate, respectively. Cell differentiation was measured using morphological investigation and cytosolic analysis of glycerol-3-phosphate dehydrogenase(GPDH; for 3T3-L1) and creatine kinase(CK; for L6). Based on the GPDH results, the presence of L6 cells did not stimulate 3T3-L1 differentiation showing more differentiation of 3T3-L1 cells in the single-culture compared to the co-culture condition. In contrast, 3T3-L1 cells in the co-culture promoted differentiation of L6 cells. Enzymatic analysis supported this result showing that 3T3-L1 cells showed statistically(P<0.05) higher GPDH activity in the single-culture than the co-culture, whereas CK results of L6 cells were vice versa(P<0.05). Overall, present results may indicate that co-culture system is more reliable and precise technique compared to single-culture. Further studies on several co-culture trials including different media conditions, supplementation of differentiating substances, molecular biological analysis, etc. should be required to obtain practical and fundamental mass data.
유전 및 육종 : 초위성체 표지를 이용한 한국재래돼지 집단의 분자유전학적 고찰
연성흠 ( Seong Heum Yeon ),이승수 ( Seung Soo Lee ),조창연 ( Chang Yeon Cho ),진현주 ( Hyun Ju Jin ),유용희 ( Yong Hee Yoo ),이풍연 ( Poong Yeon Lee ),위미순 ( Mi Soon Wee ),고응규 ( Yeoung Gyu Ko ),손준규 ( Jun Kyu Son ) 한국동물자원과학회(구 한국축산학회) 2011 한국축산학회지 Vol.53 No.1
초위성체(MS) 표지를 이용하여 한국재래돼지 집단의 각각의 분자유전학적 특성을 조사하고 그 평가를 통해 한국재래돼지에 대한 품종 및 계통분류의 기초를 마련하고자 본 연구를 수행하였다. 또한, 각 재래돼지 집단 내 및 집단간의 유전적 변이성을 확인하고, 그 분류 및 특성평가를 위한 MS 분석체계를 마련하여 국내 가축유전자원 관리에 활용하고자 하였다. 국내 관리기관 및 농가에서 보유하고 있는 6개 재래돼지 집단을 중국의 4개 재래돼지 집단 및 외래종 돼지 7개 집단과 함께 분석하였다. 도합 17집단 648두를 대상으로 26개 MS 표지로 분석한 결과, 한국재래돼지 집단은 외래종과 중국재래돼지로부터 분자유전학적으로 별개의 집단으로 확연히 구분되는 것을 확인하였다. 한국재래돼지 집단의 기대이형접합도(HE)는 0.65의 값을 보인 두 집단(B, D)을 제외한 나머지에서 0.48~0.55의 수준을 보여 전반적으로 외래종에 비해 낮았다. 한국재래돼지 집단간의 유전거리 또한 0.12~0.34 정도로 비교대상에 비하여 낮았다. 분석대상 한국재래돼지 6집단 중 세 개의 집단은 높은 유전적 균일도를 보였으나, 두 집단에서는 일부 집단의 혼입을, 나머지 하나의 집단에서는 둘 이상의 집단으로부터의 복잡한 혼입이 의심되는 매우 낮은 유전적 균일도를 확인하였다. 본 연구를 통하여 한국재래돼지 집단간의 유전적 차이 및 동질성, 그리고 집단내의 유전적 균일성을 확인하였다. 이러한 결과는 국내유전자원의 고유성을 인정할 수 있는 과학적인 근거로서, 국가수준의 가축유전자원 평가, 관리의 기초자료로 활용될 수 있을 것이다. The study was conducted to select and optimize microsatellite(MS) markers for evaluation of Korean native pig(KNP) populations in order to provide standard for the classification and breed definition of the indigenous breeds. The study also aimed to characterize and classify each KNP populations. A total of 648 pigs from 17 pig populations including six KNP, four Chinese native pig and four commercial pig populations were analyzed with 26 MS markers. KNP populations formed separate cluster from those of Chinese native pig and introduced pig populations. Expected heterozygosity(He) of KNP populations were 0.48~0.55 except two populations with 0.65. Genetic distances between KNP populations were relatively shorter: 0.12-0.34. Among six KNP populations, three showed high genetic uniformity, two showed lower uniformity and one showed high level of impurity and heterozygosity. The results can be used to evaluate and manage animal genetic resources at national scale.
Sang-Rae Cho,Kuldeep Kumar,Seong-Heum Yeon,Sung-Hwan Lee,Ui-Hyung Kim,Ki-Yong Chung,Hyeon-Shup Kim,Myeung Sik Lee,Chang-Seok Park,Byoung-Chul Yang 한국수정란이식학회 2014 한국동물생명공학회지 Vol.29 No.4
The present study was performed on farm animals to test the effectiveness of progesterone-releasing intravaginal device (Cue-Mate® 1.56 g) and injection of prostaglandin F2α (PGF2α) for synchronization estrus in Hanwoo cattle. The cattle were at random stage of the estrus cycle. The cows were artificially inseminated at day 7 after Cue-Mate withdrawal, using commercial semen from Korean native bulls. There was a season effect on the estrus synchronization rate. It was higher in spring (94.3%) followed by winter (93.3%), autumn (90.4%) and summer (67.2%). In summary, The results of this study revealed that season has influences on estrus behavior of cattle with no significant effect on pregnancy rate. In summary, we suggest summer reproductive management to alleviate the effects of heat stress. It should be based on intensive cooling combined with hormonal treatment. Given that different subgroups of cows benefit differently from the treatments, selective hormonal administration should be considered.
Seong, Eun Soo,Yoo, Ji Hye,Choi, Jae Hoo,Kim, Chang Heum,Jeon, Mi Ran,Kang, Byeong Ju,Lee, Jae Geun,Choi, Seon Kang,Ghimire, Bimal Kumar,Yu, Chang Yeon Hindawi Publishing Corporation 2015 International journal of genomics Vol.2015 No.-
<P><I>Perilla frutescens</I> is valuable as a medicinal plant as well as a natural medicine and functional food. However, comparative genomics analyses of <I>P. frutescens</I> are limited due to a lack of gene annotations and characterization. A full-length cDNA library from <I>P. frutescens</I> leaves was constructed to identify functional gene clusters and probable EST-SSR markers via analysis of 1,056 expressed sequence tags. Unigene assembly was performed using basic local alignment search tool (BLAST) homology searches and annotated Gene Ontology (GO). A total of 18 simple sequence repeats (SSRs) were designed as primer pairs. This study is the first to report comparative genomics and EST-SSR markers from <I>P. frutescens</I> will help gene discovery and provide an important source for functional genomics and molecular genetic research in this interesting medicinal plant.</P>
肉用種雄鷄의 飼養方法이 體成長 및 性成熟에 미치는 影響
延成欽,朴昌植,金榮默 충남대학교 농업과학연구소 1986 農業技術硏究報告 Vol.13 No.1
This experiment was conducted to find out the effects of the six different feeding methods on the development of body weight, testis, comb and pituitary gland, and the sexual maturity of White Plymouth Rock cockerels. From hatching to 22 weeks of age, the weights of whole body, testis, comb and pituitary gland, and the histological changes of testis and the semen characteristics were checked every other week. The results obtained in this expeniment were as follows: 1. The growth rates of the self-feeding groups were faster than those of the limited feeding groups (70 percent of the self-feeding) by about 2 weeks. The weights of testis and comb showed the most marked increase at 20 weeks of age in the self-feeding groups and at 22 weeks of age in the limited feeding groups, respectively. 2. The weights of pituitary gland from hatching to 22 weeks of age at all observation weeks were not recognized significantly among the compared groups except 4, 14 and 16 weeks of age. 3. Correlations between week of age, body weight, testis, comb and pituitary gland, in the course of 22 weeks, were highly significant. 4. The diameters of lumina and tubules in the seminiferous tubules increased very slowly until 10 weeks of age. They showed the most marked increase at 12 weeks of age in the self-feeding groups and at 14 weeks of age in the limited feeding groups, and then continueously increased until 22 weeks of age. 5. Primary spermatocytes appeared at first at 8 weeks in the all treatment groups. Secondary spermatocytes appeared at first at 10 weeks in the self-feeding groups and at 12 weeks in the limited feeding groups. At 14 weeks of age spermatids and spermatozoa were found at first in the self-feeding groups but spermatids were found in the limited feeding groups. 6. Age of the first ejaculation was between 14 and 16 weeks of age in the all treatment groups. The average semen volume and sperm concentration ranged from 0.1∼0.2 ㎖/ejaculate and 5.6∼9.8×10^8 sperm/㎖ at the age of the first ejaculation but 0.30∼0.35㎖/ejaculate and 22.4∼42.7×10^8 sperm/㎖ at the 20 weeks of age in the all treatment groups.