• 제목/요약/키워드: Culture medium

검색결과 5,722건 처리시간 0.041초

마늘의 경정배양에서 기내인경구 대량생산을 위한 2단계 배양의 도입 (Introduction of two-step culture method for multiple seed bulb development from shoot tip culture of garlic (Allium sativium L.))

  • 황혜연;이영복
    • Journal of Plant Biotechnology
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    • 제35권1호
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    • pp.75-80
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    • 2008
  • 마늘의 경정을 기내배양하여 대량의 기내종구를 생산하기 위한 다신초의 유기 및 기내구의 비대조건을 찾고자 배지의 당의 농도를 다르게 하고 또한 생장조절물질을 첨가하여 신초의 대량발생 여부를 확인하였으며, 대형배양용기를 이용하여 대량생산을 하기 위한 tank배양에서 신초의 분화와 기내구의 발달에 관하여 검토하였다. 경정배양에서의 shoot 발생이나 기내구 형성 모두 3% sucrose 첨가 MS고체배지에서 8% 보다 양호하였으며, 생장조절물질의 효과는 2 mg/L 2iP와 0.2 mg/L IAA의 혼합처리에서 shoot 발생과 bulb의 형성수가 양호하였다. 이 처리구에서 발생한 shoot와bulb의 수는 각각 3.3 및 2.7개로 나타났다. 다신초를 유도할 수 있는 조건의 배지에서 경정을 2개월 동안 배양하여 다신초를 분화시킨 후, 20 L water tank를 이용하여 sucrose 농도를 3%와 8%로 설정한 MS액체배지에 옮겨 45일간 배양한 후에 bulb형성을 조사하였을 때 유식물체의 생육에 있어서는 3% sucrose 첨가조건에서 8% 첨가배지보다 양호하였다. 그러나 기내구의 형성이나 생장에 있어서는 sucrose의 농도가 8%일 때 보다 효과적이었다. 경정배양에서의 다신초의 초기생산에는 3% sucrose조건에서 양호한 효과를 보이고 있으므로 마늘의 인경구를 대량으로 생산하기 위해서는 다신초의 생산과 구비대 과정을 구분하는 2단계 배양법을 도입하는 것이 유리할 것이다. Tank배양시 배지에 0.2 mg/L BA, 0.02 mg/L NAA를 첨가한 처리에서 양호하였으며 NAA 농도가 높을 수록 유식물체의 생육이나 구비대의 효과는 현저하게 저하되었다.

Effects of culture media conditions on production of eggs fertilized in vitro of embryos derived from ovary of high grade Hanwoo

  • Lee, Jun Young;Jung, Yun Gil;Seo, Byoung Boo
    • Journal of Animal Science and Technology
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    • 제58권3호
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    • pp.11.1-11.6
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    • 2016
  • Background: This study was investigated the effects of culture media conditions on production of eggs fertilized in vitro of embryos from ovaries of high grade Korean native cow, Hanwoo. Methods: The IVMD 101 and IVF 100 were used for in vitro maturation of selected Hanwoo oocytes and In vitro embryo culture after in vitro fertilization, respectively. The IVMD 101 and IVD 101 were used for in vitro culture and completely free of serum. Results: The cleavage rates of 2-cell embryos in reference to Hanwoo oocytes were 86.7, 92.9, and 90.1 % in the control group, IVDM101 medium and IVD101 medium, respectively which indicates that the IVDM101 medium and IVD101 medium may result favorable outcomes. The in vitro development rates of blastocysts were 12.4, 38.4 and 32.4 % in the control group, serum free IVMD101 medium and IVD101 medium, respectively. For hatched blastocysts, it was 5.3, 33.9, and 28.6 % in the control group, serum free IVMD101 medium and IVD101 medium, respectively. Hence, more favorable results were expected for the hatched blastocysts in which the IVMD101 medium and IVD101 medium were used than the control group. Average cell numbers of blastocysts were 128.3, 165.7, and 163.6 in the groups of TCM-199 + 10 % FBS medium, IVMD 101 medium, and IVD 101 medium, respectively which clearly show that the IVMD 101 and IVD 101 medium consequence significantly higher cell numbers compared to the control group (i.e., TCM-199 +10 % FBS medium). Pregnancy rate after embryo transfer was 39.6 % when the serum free medium was used which is higher than that of the medium supplemented with serum (32.8 %). In addition, stillbirth rates were 4.9 % in the group of serum free medium whereas it was 13.6 % in the serum supplemented medium (13.6 %). Conclusions: Taken altogether, serum free media, the IVMD 101 and IVD 101 represented more favorable results in the embryo development rate of embryos, cell numbers of blastocyst, and pregnancy rate. Of note, the IVMD 101 medium showed better outcomes hence, it might be a better option for future applications for in vitro culture of bovine embryos.

양배추 배축 원형질체로부터 식물체 재분화 (Plant Regeneration from Hypocotyl-Derived Protoplasts of Brassica oleracea var. capitata)

  • 이연희;조현석;서석철;김호일
    • 식물조직배양학회지
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    • 제22권1호
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    • pp.7-11
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    • 1995
  • 종자발아 후 5일된 양배추 그린챌린져 배축으로부터 분리된 원형질체로부터 완전한 식물체를 재분화 시켰다. Kao배지와 B5 배지를 기본으로 한 기존의 몇가지 배지에서 원형질체를 배양한 결과 변형시킨 BS배지에서 Plating efficiency는 낮았으나 식물체 재분화율은 가장 높게 나타나 거의 원형질체 배양배지 조성, 생장조절제 농도 등이 재분화에 상당한 영향을 주는 것으로 나타났다. 또한 사용된25가지 종류의 재분화 배지중에서 MS배지에 l% sucrose zeatin 3 mg/L, 1.6% T.C. agar가 첨가된 배지에서 재분화율이 가장 좋았다. 식물체 재분화에 미치는 PVP (polyvinyl Polypyrrolidone)와 agar 농도의 영향으로는 PVP는 큰 효과가 없었고 agar농도를 1.6%로 2배 증가시켰을 때 재분화율이 다소 높아졌다. 현재까지 재분화된 식물체는 150여 개체로서 포장에서 재배중에 있다.

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유청단백질농축물을 기본 배지로 한 Bacillus polyfermenticus SCD균의 생육과 항산화물질 생산 (Growth and Antioxidant Production of Bacillus polyfermenticus SCD in Whey Protein Concentrate (WPC)-based Medium)

  • 최귀헌;이장현;조미나;윤여창;백현동
    • 한국축산식품학회지
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    • 제28권1호
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    • pp.105-108
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    • 2008
  • The cell growth and antioxidant activity of Bacillus polyfermenticus SCD were studied in tryptic soy broth (TSB) medium and whey protein concentrate (WPC)-based medium. Overall, higher lactose contents in WPC-35 medium (up to 2.0%), and longer culture times correlated with greater cell viability. In WPC-35 medium with 1.5% and 2.0% lactose, the cell growth of B. polyfermenticus SCD was similar to growth in TSB medium. The 1,1-diphenyl-2-picyrylhydrazyl (DPPH) radical scavenging activity of culture supernatant of B. polyfermenticus SCD in WPC-35 medium was measured to assess antioxidant activity. The antioxidant activity increased up to 32 hr of culture, reaching a maximum of 75.57% DPPH radical scavenging activity. The antioxidant activity seemed to follow the typical kinetics of primary metabolite synthesis. The antioxidant activity of B. polyfermenticus SCD supernatant in WPC-35 medium was more effective and stable than supernatant from TSB medium. These results suggest that WPC-35 medium is effective for the production of antioxidant by B. polyfermenticus SCD.

Effects of Serum Addition and Different Culture Media on Growth of Porcine Preantral Follicles In Vitro

  • Diao, Yun-Fei;Kim, Hong-Rye;Han, Rong-Xun;Kim, Myung-Yoon;Park, Chang-Sik;Jin, Dong-Il
    • Reproductive and Developmental Biology
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    • 제34권3호
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    • pp.207-211
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    • 2010
  • Current developments in IVF and animal cloning have resulted in increasing demand for large quantities of oocytes and ovarian follicles at specific stages of development. These medical and scientific needs may be met by developing an optimal culture system for preantral follicles. In this study, we investigated the growth of porcine preantral follicle cultures in different media and in the presence and absence of serum. Follicles were manually dissected from ovaries obtained from prepubertal gilts at a local slaughterhouse, and cultured for 3 days in M199 or NCSU23 medium supplemented with porcine FSH, transferrin, L-ascorbic acid and insulin. Follicle diameters were measured on day 1 and 3 of culture. In Experiment 1, the effect of supplementing culture medium with fetal calf serum (FCS) on porcine preantral follicle growth was examined. In the group of cultures supplemented with FCS, follicle diameter after 3 days of culture, survival rate and antrum formation rate in the FCS group were significantly higher than those of the control group. In Experiment 2, the effects of culture medium (M199 and NCSU23) on follicle growth were compared. Follicle diameters were increased in the M199 group, compared with those in NCSU23 (p<0.05), but we observed no significant differences in survival and antrum formation rates between cultures grown in the two media. In conclusion, supplementation of the culture medium with serum enhances preantral follicle growth and antrum formation, and M199 is superior to NUSU23 for porcine preantral follicle culture in vitro.

In vitro Culture Conditions for the Mouse Preantral Follicles Isolated by Enzyme Treatment

  • Kim, Dong-Hoon;Seong, Hwan-Hoo;Lee, Ho-Joon
    • Asian-Australasian Journal of Animal Sciences
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    • 제21권4호
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    • pp.532-537
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    • 2008
  • In order to investigate the factors affecting the culture of mouse preantral follicles in vitro, we examined the effect of culture media, protein supplements, and culture period on their growth. The oocyte diameter (initial size: $55.6{\pm}2.5{\mu}m$) was progressively increased during culture, and the maximum size ($72.0{\pm}2.4{\mu}m$) was reached on day 10 of the in vitro culture. The chromatin configuration in the germinal vesicle (GV) oocyte progressively shifted from a non-surrounded nucleolus (NSN) to a surrounded nucleolus (SN). On day 10 of the culture, most of the oocytes progressed to the SN pattern. The survival and metaphase II rates of the oocytes in alpha-minimal essential medium (alpha-MEM) were significantly higher (p<0.05) than those in Waymouth and tissue culture medium (TCM)-199. As a protein source, fetal bovine serum (FBS) was more suitable for the culture of mouse preantral follicles as compared to human follicular fluid (hFF) and bovine serum albumin (BSA); the optimal concentration of FBS was 5%. These results suggest that in a culture of mouse preantral follicles, alpha-MEM and 5% FBS are an optimal medium and a protein source, respectively; further, the 10 days of culture is required for the complete growth of oocytes in this culture system.

High-Cell-Density Fed-Batch Culture of Saccharomyces cerevisiae KV-25 Using Molasses and Corn Steep Liquor

  • Vu, Van Hanh;Kim, Keun
    • Journal of Microbiology and Biotechnology
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    • 제19권12호
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    • pp.1603-1611
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    • 2009
  • High-cell-density cultivation of yeast was investigated using the agricultural waste products corn steep liquor (CSL) and molasses. The Saccharomyces cerevisiae KV-25 cell mass was significantly dependent on the ratio between C and N sources. The concentrations of molasses and CSL in the culture medium were statistically optimized at 10.25% (v/v) and 16.87% (v/v), respectively, by response surface methodology (RSM). Batch culture in a 5-l stirred tank reactor using the optimized medium resulted in a cell mass production of 36.5 g/l. In the fed-batch culture, the feed phase was preceded by a batch phase using the optimized medium, and a very high dried-cell-mass yield of 187.63 g/l was successfully attained by feeding a mixture of 20% (v/v) molasses and 80% (v/v) CSL at a rate of 22 ml/h. In this system, the production of cell mass depended mainly on the agitation speed, the composition of the feed medium, and the glucose level in the medium, but only slightly on the aeration rate.

느타리버섯 재배용 배지 입상 장치 개발(2) - 시작기 성능시험 및 경제성 평가 - (Development of Loading Machine of Culture Medium for Oyster Mushroom Production - Performance Test and Economic Analysis of Loading System -)

  • 이경진;임학규;김태한
    • Journal of Biosystems Engineering
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    • 제34권4호
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    • pp.220-227
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    • 2009
  • In the process of oyster mushroom production, loading work of culture medium needs the most intensive labor power. Therefore, development of culture medium machine causes to reduce the manpower and cost. The main objective of this study is to develop the culture medium loading machine and investigate the optimal operation conditions and to evaluate the economic value of the machine. The results are summarized as follows: 1. Optimum transporting velocity of the conveyor was 0.61 m/s 2. Optimum speed of blower was 3183 rpm at the transporting velocity of 0.61 m/s with the loading quantity of 3.41 t/hr 3. Recommendable opening area ratio of pressure controller was 1/2 at the blower speed of 3183 rpm and the transporting velocity of 0.61 m/s 4. The break even point resulted in $240\;m^2$ of cultivating area compared to the method of with portable workbench, and $350\;m^2$ of cultivating area compared to the method of with a tractor and a truck.

난관상피세포 Conditioned Medium이 체외수정된 소 수정란의 체외 발달에 미치는 영향 (Effect of Bovine Oviductal Epithelial Cell(BOEC) Conditioned Medium on In Vitro Development of Bovine Embryos Fertilized In Vitro)

  • 오종훈;김동훈;정형민;이훈택;정길생
    • 한국가축번식학회지
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    • 제17권1호
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    • pp.69-74
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    • 1993
  • This study was investigated to examine the effect of conditioned medium from bovine oviductal cell(BOEC) in the co-culture system with BOEC on in vitro development of in vitro produced bovine embryos. Oocyte-cumulus complexes were cultured for 24 hrs in TCM-199 supplemented with 10% fetal calf serum, 1$\mu\textrm{g}$/ml FSH and 21U hCG, 1$\mu\textrm{g}$/ml oestradiol-17$\beta$ at 39$^{\circ}C$ under 5% CO2 in air. In vitro fertilization was performed with epididymal sperm and heparin (10$\mu\textrm{g}$/ml, 15min.) or caffeine(2.5mM)-treated spermatozoa. Oocytes were incubated with 1$\times$106 spermatozoa/ml for 18 hrs and then cultured in various culture system for 7 days. The development rates of 16-cell or blastocyst stages were recorded on 4, 7 days, respectively, after incubating. The proportions ofembryonic development into molulae and blastocysts were higher in cumulus cell co-culture(23.4%) and BOEC co-culture(34.3%) than in M199-FCS(6.1%). Similarily, the development rates into molulae and blastocysts were significantly higher in BOEC-conditioned medium than those in M199-FCS. Therefore, it is suggested that BOEC co-culture and BOEC conditioned medium increase significantly the development of in vitro produced bovine embryos in in vitro system.

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사과겹무늬썩음병균(病菌) Botryosphaeria dothidea에 의한 Pectin질(質) 분해효소(分解酵素)의 생산 (Production of Pectolytic Enzymes by Botryosphaeria dothidea)

  • 박석희;김기홍;이창은
    • 한국균학회지
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    • 제19권2호
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    • pp.143-147
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    • 1991
  • 사과겹무늬썩음병균 Botryosphaeria dothidea를 pectin-polypectate 배지(培地)와 사과배지(培地)에 배양(培養)하여 pectin질(質) 분해효소(分解酵素)의 생산(生産)과 활성(活性)을 조사하였다. exo-polygalacturonase와 exo-polymethylgalacturonase의 활성은 사과배지에서 배양 6일에 각각 6.4 와 7.2 units로 최대의 활성을 나타내었고, 인공배지에서는 배양 8일에 각각 5.9 와 5.3 units 의 활성을 나타내었으나 사과배지에 비해 낮았다. endo-polygalactu-ronase와 endo- polymethylgalacturonase는 인공배지에서 각각 4.4 와 16.2 units로 배양 8 일과 10일에서 최대활성을 나타내었으나 사과배지에서는 배양 8일에 각각 3.2와 6.7 units로 낮은 활성을 보였다. pecti-nmethyl-trans-eliminase와 polygalacturonate-trans-eliminase의 활성은 사과배지보다 인공배지에서 높게 나타났으며, 균 생장은 인공배지와 사과배지에서 각각 배양 6일과 8일에 최고의 생장을 나타내었다.

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