• 제목/요약/키워드: culture-dependent

검색결과 1,198건 처리시간 0.03초

Effect of Co-Culture with Mammalian Spermatozoa on In Vitro Maturation of Porcine Cumulus-Enclosed Germinal Vesicle Oocytes

  • Kim Byung Ki
    • Reproductive and Developmental Biology
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    • 제28권4호
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    • pp.235-240
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    • 2004
  • In vitro maturation of denuded porcine immature oocytes can be enhanced by co-incubation with spermatozoa even before fertilization. This study was to determine whether the addition of spermatozoa into the culture medium could influence the nuclear maturation of porcine cumulus-enclosed germinal vesicle (GV) oocytes. Cumulus-oocyte complexes (COCs) were collected from follicles of 3- to 5-mm diameter. Porcine COCs were cultured in tissue culture medium containing spermatozoa. After 48 h culture, oocytes were examined for evidence of GV breakdown, metaphase I, anaphase-telophase I, and metaphase II. The proportion of oocytes reaching at metaphase II was significantly (P < 0.05) increased in the oocytes cultured in media containing spermatozoa compared to those in media without spermatozoa (52.3% vs 12.5%). No difference in the percentage of metaphase II was observed among the different periods of spermatozoa exposure and among the spermatozoa from different species. The proportion of oocytes reaching metaphase II was significantly different between high and low concentrations of spermatozoa. The present study suggests that manunalian spermatozoa contain a substance(s) that improves nuclear in vitro maturation of porcine cumulus-enclosed GV oocytes. Enhancing effect of spermatozoa for in vitro maturation of oocytes is a highly dose-dependent.

미혼성인자녀가 지각하는 자아분화와 부모-자녀간 정서체계 역동성이 특성불안에 미치는 영향 (The Effects of Premarital Adult Child's Perceived Self-Differentiation and Dynamics of Emotional System between Parent and Child on Trait Anxiety)

  • 권미애;김필숙;박지현;김태현
    • 대한가정학회지
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    • 제43권11호
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    • pp.31-48
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    • 2005
  • The purpose of this study was to explore the direct and indirect effects of premarital adult child's perceived self-differentiation and parent-child dynamics of emotional system on trait anxiety. The subjects of the study numbered 277, and correlation and path analysis was used with SPSS 10.0. The major findings were as follows: 1. It was shown that self-differentiation, over involvement and perceived criticism of premarital adult children were correlated with trait anxiety. And, among demographic variables, education, income, and marital status of the subjects' parents were related with trait anxiety. 2. Parent-child differentiation, over involvement, perceived criticism, and sex had direct effects on premarital adult child's self-differentiation. The variables that had direct effects on trait anxiety, which was the final dependent variable were self-differentiation, parent-child differentiation, perceived criticism, and income while over involvement an indirect effect on it.

Effect of Epididymal Fluid on In Vitro Maturation and Subsequent Sperm Penetration in Porcine Follicular Oocytes

  • Kim, Byung-Ki
    • Reproductive and Developmental Biology
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    • 제33권3호
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    • pp.125-131
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    • 2009
  • The aim of this study is to investigate the effect of porcine epididymal fluid (pEF) on in vitro-maturation and subsequent fertilization of porcine follicular oocytes. Porcine cumulus-oocytes complexes retrieved from antral follicles were cultured in tissue culture medium (TCM)-l99 supplemented with pEF of different concentrations. At 48 h after culture, development of oocytes to germinal vesicle (GV) breakdown, metaphase I, anaphase-telophase I, and metaphase II were examined Significant (p<0.05) increase in the proportion of oocytes developed to MII stage was observed in oocytes cultured in pEF-containing TCM-l99 than in oocytes cultured in pEF-free TCM-199 (46.2% vs 16.7%), which was a dose-dependent manner. Subsequently, the proportion of monospermic fertilization were significantly (p<0.05) increased in oocytes cultured in the TCM supplemented with pEF than those cultured in pEF-free TCM-199 (51.0% vs 24.1%). In the second series of experiment, the percentage of MII oocytes was significantly (p<0.05) increased after exposure of oocytes to pEF during the first 22 h period of culture than after exposure of oocytes to pEF during the next 24 h of culture, while no significant difference in the percentage of monospermy was observed. The results of this study demonstrate that pEF contains at least enhancing component(s) for nuclear maturation.

인삼(人蔘) Saponin이 Prolactin 분비(分泌)에 미치는 효과(效果) (The Effects of Ginseng Saponin on Prolactin Secretion in Rats)

  • 백덕우;이서윤;지형준
    • 생약학회지
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    • 제9권1호
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    • pp.33-39
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    • 1978
  • The present study is involved with the prolactin secretion from anterior pituitary gland by ginseng saponin since it was handled down by tradition that ginseng might influence the milk secretion when it was given to nursing mother. To investigate the effect of saponin on the prolactin production or release from the anterior pituitary gland, cell culture study and whole animal studies were carried out. For the cell culture study, enzymatically dispersed anterior pituitary cells of rat anterior pituitary gland in HEPES buffers containing trypsin were used. Ginseng saponin was added to the culture media and the amount of prolactin produced in the cell culture media was determined by radloimmunoassay(RIA) technique. Dose-dependent increases of prolactin with ginseng saponin were observed, whereas, no change was observed without ginseng treatment. For the whole animal study, normal and castrated rats which previously cannulated into the heart via the right juglar vein were used. The prolactin concentration in plasma were determined by using the technique of RIA. In normal rats, prolactin concentration in plasma were elevated dramatically after 1 hour of ginseng saponin administration, whereas, instantaneous increases were observed in castrated rats. For prolactin assay by RIA, NIAMDD Rat Prolactin Kit and NIAMDD Rat Prolactin RP-1 were used as standard. The results indicate that ginseng saponins increase the release of prolactin from the anterior pituitary gland and production of prolactin from the cell in rats.

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가구 특성에 따른 김치 소비량 차이에 관한 연구 (A Study on the Differences of Kimchi Consumption according to Household Characteristics)

  • 박성훈
    • 한국식생활문화학회지
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    • 제34권2호
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    • pp.159-167
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    • 2019
  • The purpose of this study is to provide basic information to improve understanding of contemporary kimchi culture in Korea. Many Koreans are now purchasing kimchi at markets, while the proportion of self-preparation is gradually decreasing. This commodification tendency of kimchi is considered to be associated with changes in consumer's behavior and attitude. In this study, a linear regression and a logistic regression model were used to identify relationships between kimchi consumption behavior and household characteristics. The results showed that the probability of kimjang activity was positively related with family size, possession of a kimchi refrigerator, self-preparation practice, and the intensity of sharing behavior. I also found that kimchi consumption volume per capita of 'purchasing' household was greater than that of 'self-preparing ' or 'sharing-dependent' households, and that the number of family members was inversely related with kimchi consumption volume per capita. The inverse relationship between family size and kimchi consumption volume per capita is considered to be contrary to the widespread thoughts in Korea, which have been developed while experiencing kimchi preparation and consumption in traditional extended families. I think that the relationship comes from differences in menu varieties, which appear to vary with family size. This issue will be investigated in subsequent studies.

HaCaT 세포에서 Lactobacillus 혼합배양액 추출물이 아토피관련 케모카인 발현에 미치는 효과 (The Effect of Lactobacillus Mixture Culture Fluid Extracts on Atopic Dermatitis Chemokine Expression of in HaCaT Cells)

  • 홍수정;이원재;조을화;안성훈
    • Korean Journal of Acupuncture
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    • 제34권2호
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    • pp.82-87
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    • 2017
  • Objectives : Recently the case of lactobacillus mixture culture fluid appliment was reported. In this study, anti-inflamation effects and anti-allergy effects were studied by stimulus of lactobacillus mixture culture fluid extracts in HaCaT cells. Methods : The atopic dermatitis were induced by TNF-${\alpha}$ and interferon-${\gamma}$ in HaCaT cells. TARC/CCL17, MDC/CCL22, RANTES/CCL5 and ROS production were investigated to explain anti-inflamation and allergy effects of lactobacillus mixture culture fluid with cell-enzyme-linked Immunosorbent assay in 450 nm, 485 nm, 535 nm with spectro-fluorometer. Results : The extracts of lactobacillus mixture culture fluid were decreased TARC/CCL17, MDC/CCL22, RANTES/CCL5 expressions and ROS production with a concentration dependent manner. Conclusions : The effects mechanism of Lactobacillus mixed culture fluid for atopic dermatitis symptoms were considered to be explain anti-inflamation and allergy effects via control of cytokine, chemokine and ROS production, and the fluid could be applied in skin cells directly. But classified AD symptom degrees reported in clinical case before as Reaction Period, Reduction Period, Effect Period, Reproduction Period and Rebound Period could not be explained. Further study will be expected.

키토산이 치은섬유아세포에 미치는 영향 (Effects of Chitosan on Human Gingival Fibroblasts in Vitro)

  • 김옥수;정현주
    • Journal of Periodontal and Implant Science
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    • 제32권1호
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    • pp.235-247
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    • 2002
  • The aim of this study was to evaluate the effects of chitosan coating on the attachment, proliferation, functional and morphological change of human gingival fibroblasts. Primary culture of human gingival fibroblasts were grown in Dulbecco's modified Eagle's medium with 10% fetal bovine serum and 1% antibiotics. In experimental group, cells were inoculated in the multiwell plates coated with chitosan in concentration of 0.02, 0.2, and 2 mg/ml. Cell counting and MTT assay were done after 0.5, 1.5, 3, 6 and 24 hours of incubation to evaluate the cell attachment, and then after 2 and 7 days of culture to evaluate the cell proliferation. The alkaline phosphatase activity was measured after 4 and 7 days of culture and the ability to produce mineralized nodules was evaluated after 21 days of culture. The results were as follows : The morphology of cells on the chitosan-coated well was round or spheric. Round cells were aggregated since 6 hours of culture and showed nodule-like appearance after 24 hours of culture and did not achieved confluency at 7 days. The attachment of gingival fibroblasts was inhibited by chitosan coating with a tendency of dose dependent pattern. But, cellular activity of unit cell was higher than control. The proliferation of gingival fibroblasts was inhibited by chitosan coating at 2 mg/ml(P<0.01), while the cell proliferation at 0.02, 0.2 $mg/m{\ell}$ was comparable to the control well. Total alkaline phosphatase activity was inhibited by chitosan coating and decreased in the course of time. While ALP activity of unit cell was the highest at 2mg/ml after 4 days of culture. Finally, gingival fibroblasts produced the mineralized nodule at 2 mg/ml. In summary, the attachment, proliferation, and alkaline phosphatase activity of gingival fibroblasts were influenced differently by the concentration of coated chitosan. From this study, it could be used as the matrix of tissue engineering for gingiva without inhibition on proliferation of gingival fibroblasts using chitosan at the optimal concentration (0.02mg/ml).

베큘로 바이러스 발현 시스템에 의한 곤충세포에서의 인간 트롬보포이에틴 생산 최적화 (Optimization of Human Thrombopoietin Production in Insert Cells Using Baculovirus Expression System)

  • 고여욱;손미영;박상규;안혜경;박승국;박명환;양재명
    • KSBB Journal
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    • 제13권2호
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    • pp.181-186
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    • 1998
  • 부착의존성 세포주인 Trichoplusia ni 의 유래의 BTI-TN5B1-4 (TN5) 곤충세포주를 이용하여 인간 혈소판생성축진인자인 재조합 인간 트롬보포이에틴(rhTPO)의 배양조건 최적화 연구를 수행하였다. 배양배지, 세포감염에 투입되는 재조합 베큘로바이러스와 숙주세포의 비율(MOI),세포감염시 세포밀도, 배지 회수시간 및 배양방법 등이 rhTPO 의 생산에 미치는 효과를 연구하여 60 mm dish로 정체 배양시 10 MOl 이상,$2\times10^6$ cells 의 세포밀도,바이러스 감염 후72 시간에서 rhTP0 의 최대 발현양 (약 12 mg/L)을 나타내었다. 배양 배지로서는 EXCELL FIVE 배지가 SF900II나Insect serum free media-1 Figure 5. Effect of growth phases on rhTPO production. TN5 cells were grown as suspension culture in 1 L spinner flask with 200 mL of SF900II serum free medium at 80 rpm. The cells were infected with AcBac404-2 at MOl of 1. Culture medium was collected at given time intervals and the expression level of rhTPO was analyzed by ELISA (A) or immunoblot analysis (B). Lanes 1 and 7; cell density of $0.6\times10^6$ cells/mL, lanes 2 and 8; cell density of $1.6\times10^6$ cells/mL, lanes 3 and 9; cell density of $2.0\times10^6$ cells/mL, lanes 4 and 10; cell density of $3.0\times10^6$ cells/mL, lane M; prestained molecular weight marker (Bio-Rad). Lanes 1, 2, 3, and 4; culture medium was collected at 48 hpi and lanes 7, 8, 9, and 10; culture medium was collected at 72 hpi. Figure 6. Effect of culture media on rhTPO production. TN5 cells grown with different culture media were infected with AcBac-404-2 at 10 MOL 10$\mu$L of culture medium was run on SDS-PAGE and Immunoblot analysis was performed. Lane ];TN5 cells cultured with SF900II serum free media(Gibco),and lane 3; TN5 cells cultured with EXPRESS FIVE serum free media (Gibco) 에 비해 더 증가된 발현양을 나타내었다. TN5 세포주를 0.2 L 규모 (1 L spinner flask)oJl에서 세포간의 응집현상 없이 부유배양에 적응,배양시킨 후 세포성장 시기에 따른 발현을 조사한 결과 1 MOI의 감염조건 하에서는 $0.6\times10^6$cell/mL의 early exponential시기의 세포밀도에서 72시간 배양하였을 대 최대 발현양을 나타내었다. 나타내었다.

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비살균 숙성 치즈의 미생물균총 분석에 이용되는 새롭게 개발된 분자생물학적 방법: 총설 (Novel Molecular-Based Approaches for Analyzing Microbial Diversity in Raw-Milk Long-Ripened Cheeses: A Review)

  • 김동현;천정환;김현숙;이수경;김홍석;이주연;임진혁;송광영;김영지;강일병;정다나;박진형;장호석;서건호
    • Journal of Dairy Science and Biotechnology
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    • 제34권1호
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    • pp.9-20
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    • 2016
  • Various microflora, including lactic acid bacteria, are important and necessary components of various cheeses and have significant roles in cheese manufacturing and ripening. In general, the starter culture and secondary microflora could affect the physicochemical properties of various cheeses and could contribute to modifications during manufacturing and ripening. Therefore, during cheese manufacturing and ripening, microbial diversity may depend on continuous interactions among microflora and various environmental conditions. The microbial diversity of cheese is very complex and difficult to control using the classical microbiological techniques. However, recent culture-independent methods have been rapidly developed for microflora in cheese, which could be directly detected using DNA (and/or RNA) in combination with culture-dependent methods. Therefore, this review summarizes state-of-the-art molecular methods to analyze microbial communities in order to understand the properties that affect quality and ripening as well as the complex microbial diversity of various raw-milk, long-ripened cheeses.

새로운 생균제 CS61 배양액의 사료 내 급여가 육계의 생산성에 미치는 영향 (Effects of Dietary Supplementation of a New Probiotic CS61 Culture on Performance in Broiler Chickens)

  • 김성환;이인철;백형선;강성수;김형진;유진철;김종춘
    • 생명과학회지
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    • 제22권3호
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    • pp.340-346
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    • 2012
  • 가금류 제품에서 항생제의 잔류와 내성균의 출현으로 인해 항생제 대체물질에 대한 연구가 활발히 진행되고 있으며, 특히 생균제와 활성촉진제 또는 이들을 조합한 신바이오틱의 사용이 권장되고 있다. 본 연구는 새로운 생균제 CS61 배양액의 사료첨가 급여가 육계의 성장 및 사료효율에 미치는 영향과 안전성을 평가하여 항생제를 대체할 수 있는 사료첨가제로서의 개발가능성을 알아보기 위해 수행하였다. CS61 배양액은 0, 0.1 및 1%의 용량으로 28일간 사료에 혼합하여 육계에게 급여하였다. 시험결과, 시험물질 처치군에서 부검 시의 체중과 일당증체량이 대조군에 비해 용량의존적으로 증가하였다. CS61 배양액의 사료 내 첨가는 대조군 동물에 비해 사료효율도 개선하는 것으로 나타났다. 반면, 일반증상과 사망률, 부검소견, 혈액학치 및 혈청생화학치에서는 시험물질의 처치와 관련된 독성소견이 관찰되지 않았다. RAW 264.7 세포를 이용한 일산화질소 시험에서 정제된 CS61 펩타이드는 lipopolysaccharide에 유도된 일산화질소 생성을 용량의존적으로 억제하였다. 본 시험결과는 육계에 CS61 배양액의 사료첨가 급여는 항염증효과를 통해 성장과 사료효율을 개선할 수 있음을 보여주며, 사료첨가제로서 CS61 배양액의 유용성과 개발가능성을 시사해 주고 있다.