• 제목/요약/키워드: Germ

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Hoechst 33258 Staining을 이용한 웅성 생쥐 성세포의 간편 분류 (Simple Classification of Male Mouse Germ Cells using Hoechst 33258 Staining)

  • 김경국;박영식
    • 한국수정란이식학회지
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    • 제30권3호
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    • pp.213-218
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    • 2015
  • In the study for a differentiation and development of spermatogonial cells, the researchers should commonly require a simple, fast and reasonable method that could evaluate the developmental stage of male germ cells without any damage and also relentlessly culture them so far as a cell stage aiming at experimental applications. For developing the efficient method to identify the stage of sperm cells, the morphological characteristics of sperm cells were investigated by staining the cells with blue fluorescent dye Hoechst 33258, and a criterion for male germ cell classification was elicited from results of the previous investigation, then the efficiency of the criterion was verified by applying it to assort the germ cells recovered from male mice in age from 6 to 35 days. As morphological characteristics, spermatogonia significantly differed from spermatocytes in size, appearance and fluorescent patches of nucleus, and spermatids could also be distinguished from spermatozoa by making a difference in the volume and shape of nucleus and the shape and fluorescence of tail. Aforesaid criterion was applicable for classifying in vitro cultured sperm cells by verifying its efficiency and propriety for assorting the stages of testicular germ cells. However, the fluorescent staining showed that germ cells in mouse testis should be dramatically differentiated and developed at 21 days and 35 days of age, which were known as times of sexual puberty and maturity in male mice, respectively. In conclusion, the results indicated that this simple criterion for sperm cell classification using fluorescence staining with Hoechst 33258 may be highly efficient and reasonable for spermatogenesis study.

마우스 수컷 생식세포에서 비스페놀 A에 대한 인삼 에탄올 추출물의 보호 효과 (Protective Effect of Panax ginseng Ethanol Extracts Against Bisphenol A (BPA) in Mouse Male Germ Cells)

  • 김형돈;손상현;김진성;이희정;박춘근;안영섭;이상원;김영옥
    • 한국약용작물학회지
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    • 제23권2호
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    • pp.138-143
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    • 2015
  • This study was carried out to evaluate the preventive effect of three forms of Korean ginseng roots (fresh, white and red) against bisphenol A (BPA) toxicity in mouse male germ cells (GC-2spd, TM3, TM4). ROS (reactive oxygen species) generation were measured by DCF-DA (2',7'-dichlorohydrofluorescein diacetate) assay. Also, semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR) was performed to quantify the mRNA expression levels of apoptosis-related genes, Bax (pro-apoptotic gene) and Bcl2 (anti-apoptotic gene). ROS generation was increased by $50{\mu}M$ BPA, but definitely decreased by treatment with Korean ginseng extracts (fresh, white and red) in mouse male germ cells. In especial, Korean fresh ginseng extract reduced significantly ROS production to normal control. In addition, Korean fresh and white ginseng extracts suppressed the apoptosis of mouse male germ cells by fine-tuning mRNA levels of apoptotic genes changed by BPA. In general, Korean fresh ginseng extract was more effective than white ginseng extract for reducing BPA-induced oxidative stress and apoptosis in mouse male germ cells. Therefore, Korean fresh and white ginseng may help to alleviate biphenol A toxicity in mouse male germ cells.

국산 쌀배아의 이화학적 성분 (The Chemical Components of Korean Rice Germ)

  • 최원균;윤승길;황성연
    • 한국식생활문화학회지
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    • 제15권4호
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    • pp.253-258
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    • 2000
  • 국내에서 버려지는 쌀눈활용을 위한 기초연구로 쌀눈의 화삭성분들을 분석하였다. 일반성분은 수분 8.69%, 조지방 21.18%, 조단백질 16.50%, 조섬유 2.95%, 조회분 6.23%, 그리고 당질함량은 44.45%로 나타났다. 유리당은 maltose, glucose, fructose가 있었으며 각각 569.00, 68.80, 50.20mg/100g함유되어 있었다. 아미노산함량은 글루탐산이 1920.09mg/100g, 아르기닌이 15.03mg/100g, 아스파르트산이 1208.7mg/100g, 알라닌이 1001.2mg/100g으로 가장 많이 함유되어 있었으며, 함황아미노산인 시스틴과 메티오닌은 적었다. 무기질 조성은 8종류의 무기성분들이 검출되었으며 칼륨과 인이 각각 가장 많은 1217.80, 1766.22mg/100g이었으며 칼슘, 망감, 나트륨이 각각 36.60, 16.74, 16.22mg/100g으로 많았으며, 아연, 철, 구리도 각각 11.75, 7.99, 1.16mg/100g이 들어 이었다. 비타민함량은 5가지의 수용성 비타민과 지용성비타민이 토코페롤이 함유되어 있었으며, 비타민 E가 11.96mg/100g으로 가장 많았고 $B_1$$B_2$는 각각 5.69mg/100g, 0.45mg/100g였으며 비타민 C는 1.66mg/100g 그리고 나이아신과 비타민 $B_6$는 각각 2.96mg/100g, 0.69mg/100g였다. 지방산은 6가지 종류의 지방산이 함유되어 있었으며, 주지방산은 linoleic acid와 oleic acid로 전체지방산의 약 64%를 차지하고 있다. 또한 모든 유지류에 널리 분포되어 있는 palmitic acid가 22.2%였으며, arachidic acid도 3.1%였다. 16종의 flavonoid와 9종의 phenolic acid를 screening한 결과 쌀눈에서는 검출되지 않았다.

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밀배아유 원료 O/W 유화액의 제조 및 안정성평가: CCD-RSM을 이용한 최적화 (Emulsification and Stability of Wheat Germ Oil in Water Emulsions: Optimization using CCD-RSM)

  • 홍세흠;장현식;이승범
    • 공업화학
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    • 제32권5호
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    • pp.562-568
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    • 2021
  • 천연 밀배아유와 복합 sugar ester를 이용하여 밀배아유 원료 O/W (oil in water) 유화액을 제조하였다. HLB 값, 유화제의 첨가량 및 유화시간이 O/W wheat germ oil 유화액의 평균 입자크기, 유화점도 및 ESI에 미치는 영향을 조사하였으며, 반응표면분석방법인 중심합성계획모델에 의해 제조된 유화과정의 파라미터를 모의하고 최적화하였다. O/W wheat germ oil 유화액의 제조를 위한 최적의 공정조건은 HLB 값(hydrophile-lipophile balance value)은 8.4, 유화제의 첨가량은 6.4 wt%, 유화시간은 25.4 min이며, 최적조건에서 중심합성계획(central composite design, CCD-RSM) 모델을 통한 예측 반응치는 7일 후의 유화액을 기준으로 MDS (mean droplet size)= 206 nm, 점도 = 8125 cP, ESI 98.2%이었다. 또한 실제 실험을 통해 얻어진 유화액의 MDS, 점도 및 ESI는 각각 209 nm, 7974 cP 및 98.7%로 나타났다. 따라서 중심합성계획 모델을 통해 밀배아유 원료 유화액의 안정성을 평가하는 최적화 과정을 설계할 수 있었다.

Transgenesis and Germ Cell Engineering in Domestic Animals

  • Lee, C.K.;Piedrahita, J.A.
    • Asian-Australasian Journal of Animal Sciences
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    • 제16권6호
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    • pp.910-927
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    • 2003
  • Transgenesis is a very powerful tool not only to help understanding the basics of life science but also to improve the efficiency of animal production. Since the first transgenic mouse was born in 1980, rapid development and wide application of this technique have been made in laboratory animals as well as in domestic animals. Although pronuclear injection is the most widely used method and nuclear transfer using somatic cells broadens the choice of making transgenic domestic animals, the demand for precise manipulation of the genome leads to the utilization of gene targeting. To make this technique possible, a pluripotent embryonic cell line such as embryonic stem (ES) cell is required to carry genetic mutation to further generations. However, ES cell, well established in mice, is not available in domestic animals even though many attempt to establish the cell line. An alternate source of pluripotent cells is embryonic germ (EG) cells derived from primordial germ cells (PGCs). To make gene targeting feasible in this cell line, a better culture system would help to minimize the unnecessary loss of cells in vitro. In this review, general methods to produce transgenic domestic animals will be mentioned. Also, it will focus on germ cell engineering and methods to improve the establishment of pluripotent embryonic cell lines in domestic animals.

Artificial gametes from stem cells

  • Moreno, Inmaculada;Miguez-Forjan, Jose Manuel;Simon, Carlos
    • Clinical and Experimental Reproductive Medicine
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    • 제42권2호
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    • pp.33-44
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    • 2015
  • The generation of artificial gametes is a real challenge for the scientific community today. In vitro development of human eggs and sperm will pave the way for the understanding of the complex process of human gametogenesis and will provide with human gametes for the study of infertility and the onset of some inherited disorders. However, the great promise of artificial gametes resides in their future application on reproductive treatments for all these people wishing to have genetically related children and for which gamete donation is now their unique option of parenthood. This is the case of infertile patients devoid of suitable gametes, same sex couples, singles and those fertile couples in a high risk of transmitting serious diseases to their progeny. In the search of the best method to obtain artificial gametes, many researchers have successfully obtained human germ cell-like cells from stem cells at different stages of differentiation. In the near future, this field will evolve to new methods providing not only viable but also functional and safe artificial germ cells. These artificial sperm and eggs should be able to recapitulate all the genetic and epigenetic processes needed for the correct gametogenesis, fertilization and embryogenesis leading to the birth of a healthy and fertile newborn.

Optimization of Isoflavone Extraction from Soy Germ - Research Note-

  • Bae, Sang-Moon;Jang, Chan-Ho;Kim, Jang-Hoon;Lim, Hyun-Ae;Kim, Joo-Ryong;Kim, Jeong-Hwan;Kim, Jong-Sang
    • Preventive Nutrition and Food Science
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    • 제10권3호
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    • pp.290-293
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    • 2005
  • Soy isoflavones have drawn much attention due to their potential to prevent breast and prostate cancers, osteoporosis, heart disease, and other postmenopausal symptoms. Soy germ is one of the richest sources of isoflavones, and thus has good potential to be used as the ingredient of health foods. This study examined the extraction rate of isoflavones from soy germ at various conditions. After the effect of extraction temperature and duration on isoflavones extraction from soy germ was examined, the optimum concentration of ethanol as extraction solvent was determined. When ethanol concentration was fixed at $60\%\;(v/v)$, the maximum isoflavone extraction was achieved at 2 hrs and $30^{\circ}C$. Among various concentrations of ethanol tested, $80\%\;(v/v)$ ethanol showed the highest extraction efficiency. In conclusion, the maximum extraction of isoflavones was obtained using $80\%\;(v/v)$ ethanol as a solvent, at $30^{\circ}C$ of temperature, and 2 hrs of extraction time.

쌀눈기름의 급여가 KK 당뇨 마우스의 지질대사에 미치는 영향 (Supplementary effect of the rice germ oil on Lipid Metabolism in Diabetic KK Mice)

  • 이성현;전혜경;장순옥;이연숙
    • 한국지역사회생활과학회지
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    • 제14권3호
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    • pp.59-65
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    • 2003
  • This study was carried out to investigate the supplementary effects of the rice germ oil compared with soy bean oil on lipid metabolism of non-insulin dependent diabetic mice. Forty diabetic KK mice were fed two kinds of experimental diets with 20% lipid from soy bean oil as a control(CO) and rice germ oil(RG) for 8 weeks, respectively. Diet intake, body weight, organs weights and lipids levels of serum, liver and feces were measured. There was no significant difference in food and water intake, body weight gain and organs weights between experimental groups. But the concentrations of serum total cholesterol and LDL-cholesterol were lower in RG group than in CO group. The hepatic total lipid and total cholesterol levels of RG group were significantly lower than those of CO group. The contents of total lipid, triglyceride and total cholesterol excreted in feces of RG group were higher than those of CO group. These results suggested that rice germ oil can reduce serum total cholesterol and LDL-cholesterol and hepatic total lipid concentration of non-insulin dependent diabetic mice compared with soybean oil due to increasing fecal lipid excretion. But we need to investigate the hypolipidemic effect of this oil by supplementary level and period.

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Induction of Apoptosis in the Testes of SD Rats After Exposure to 2-Bromopropane

  • Kim, Young-Hee;Cho, Sung-Whan;Ha, Chang-Su;Kang, Boo-Hyon
    • Toxicological Research
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    • 제17권4호
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    • pp.241-248
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    • 2001
  • Exposure to 2-Bromopropane has been known to cause degeneration of male germ cells. However the mechanism underlying this process is poorly understood. The objective of this study was to determine whether or not the exposure of male Sprague-Dawley rats to 2-BP induces apoptosis in male germ cells. Male rats(N=3 or 4 in each group) were orally administered either with the corn oil vehicle (10 ml/kg body weight) or with 2-BP (3,500 mg/kg) once a day for 3 days. The presence of apoptosis was determined by TUNEL detection in situ and by an increase in DNA fragmentation. A low spontaneous incidence of apoptosis was observed in vehicle control animals, especially in pre-meiotic germ cells of stages I-VI and stages XII-XIV the seminiferous tubules. In 2-BP exposure rats, the incidence of apoptosis markedly increased at 4 h, reached a peak at 8 h (about 7-fold over control), and then decreased rapidly to control level by 48 h after the last administration. Although apoptosis induced by 2-BP occurred in all stages of germ cells, it was most pronounced in spermatogonia and early spermatocytes in stages I-VI and stages XII-XIV. Taken together, our results suggest that apoptosis is involved in the toxicity of testicular germ cells resulting in oligospermia or azoospermia after exposure to 2-BP.

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