• Title/Summary/Keyword: Co-culture

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Ethanol Production from Raw Starch by Co-Immobilized Mixed Rhizopus japonicus and zymomonas mobilis (Rhizopus japonicus와 Zymomonas mobilis의 혼합고정화 배양계에 의한 생전분으로부터 에탄올 생산)

  • 최수철;이상원;박석규;성찬기;손봉수;성낙계
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.25 no.4
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    • pp.708-714
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    • 1996
  • Ethanol production from raw starch was performed using the co-immobilized culture system of Rhizopu japonicus and zymomonas mobilis(R-Z). Glucose Production in immobilized R. japonicus culture was 2-fold higher than that in free cell culture. Ethanol production was 1.67g/L(Yp/s, 0.094) and 6.549/L(Yp/s, 0.38) in R-Z and R-Z 24 culture system, respectively. R-Z system was modified and designated as R-Z 24 system by replacing cotton plug with silicon check valve after 24h fermentation with R-Z system. Optimal substrate concentration for ethanol production in batch culture was 5%(w/v) and ethanol concentration produced was 15.02g/L(Yp/s, 0.36). Ethanol yield(Yp/s, 0.38) in fed-batch culture of 5 times with 2%(w/v) substrate was equal to that in batch culture of 2%(w/v) substrate.

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In Vitro Development of Nuclear Transplantation Bovine Embryos Using In Vitro Fertilized Embryos of Korean Native Heifers (한우 체외수정란을 이용한 핵 이식배의 체외발달에 관한 연구)

  • 박충생;공일근;노규진;이효종;최상용
    • Korean Journal of Animal Reproduction
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    • v.18 no.2
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    • pp.113-119
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    • 1994
  • To improve nuclear transplantation(NT) efficiency and to produce a large scale genetically identical cloned calves, examined the in vitro development capacity after co-culture of bovine oviductal epithelial cells (BOEC) and granulosa cells in TCM-199 supplemented with 10% fetal calf serum (FCS) with early bovine embryos derived from in vitro matured fertilized(IVM-IVF) oocyte. In addition, the age dependence of IVM oocyte on electro-stimulation and the effective electric voltage on in ivtro development of bovine NT embryos were examined. The results obtained were summerized as follows; 1. The cleavage rates of IVM-IVF bovine embryos in co-culture with bovine oviductal epithelial cells and granulosa cells were not significantly different(P<0.05), but the developmental rate into morula and blastocyst stage were different showing 38.3 and 20.2%, respectively. 2. The activation (82.5%) and development in vitro(8.6%) into later embryo stages of the aging oocytes of 32 hours post-maturation (hpm) were significantly higher than those of 24 hpm at direct current (DC) voltage of 1.5kV/cm, 60$\mu$sec pulse duration and 1 pulse time. 3. The fusion rates of NT eggs of 32 hpm following to different DC voltages from range 0.75 to 1.5kV/cm were not differ, but the developmental rate into morula and blastocyst stages at DC voltages of 0.75 and 1.0kV/cm were higher(11.4 and 12.6%, respectively) than those of 1.5kV/cm(0%). From these results, it can be suggested the optimal culture system for in vitro culture of IVM-IVF bovine embryos is a co-culture system with BOEC in TCM-199 supplemented 10% FCS. The effective time and the DC voltage for activation, electrofusion and in vitro development of NT embryos derived from IVM-IVF bovine embryo are 32hpm and 0.75~1.0kV/cm. But to improve NT efficiency, the advanced research (cell cycle synchronization, micromanipulation, culture system, etc.) is needed.

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Effect of Acetic Acid Concentration and Mixed Culture of Lactic Acid Bacteria on Producing Bacterial Cellulose Using Gluconacetobacter sp. gel_SEA623-2 (Gluconacetobacter sp. gel_SEA623-2를 이용한 Bacterial Cellulose 생산에 초산농도 및 유산균 혼합배양이 미치는 영향)

  • Kim, Kyung min;Kim, Jihyeon;Yang, Kyong Wol
    • Korean Journal of Microbiology
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    • v.50 no.3
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    • pp.227-232
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    • 2014
  • In this study, Gluconacetobacter sp. gel_SEA623-2 isolated from citrus that produces bacterial cellulose was used to examine the effect of initial concentration of acetic acid and mixed culture inoculated with Lactobacillus plantarum KCCM 80077 on productivity of bacterial cellulose. In mixed culture added with 0.5% acetic acid, the viable cell count increased from $2.4{\times}10^6CFU/ml$ to $1.1{\times}10^7CFU/ml$ after 14 days of culture, and total acidity was about 0.3% higher than single culture added with 0.5% acetic acid, which implies that additional lactic acid was produced by L. plantarum KCCM 80077. In single culture, although bacterial cellulose productivity was higher when the initial concentrations of acetic acid were 0.0% and 0.5%, than when it was 1.0%, there was no significant difference. However, in mixed culture, adding 0.5% acetic acid resulted in dry weight of $37.83{\pm}6.81g/L$ and thickness of $10.33{\pm}0.58mm$, showing a significant difference from that of single culture added with 1% acetic acid, $28.40{\pm}1.23g/L$ and $7.50{\pm}0.50mm$ (P<0.05).

The ability of in vitro cultured bovine oviduct epithelial cells in binding and maintaining motility of bull sperm (체외배양 소 난관상피세포의 정자에 대한 결합 및 활력유지능)

  • Roh, Sang-ho;Lee, Byoung-chun;Hwang, Woo-suk
    • Korean Journal of Veterinary Research
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    • v.39 no.1
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    • pp.220-225
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    • 1999
  • The aim of these experiments was to investigate the effects of bovine oviduct epithelial cells (OEC) derived from different segments to bind sperm binding and maintain their motility in vitro. In experiment 1, the number of sperm attached to OEC derived from isthmus or ampulla, the motility of unattached sperm during co-culture and fertilizing ability were assessed. In experiment 2, heparin treated sperm (hsp) or no treated sperm (nsp) were used to evaluate OEC binding ability of capacitated sperm. In experiment 1, regardless of their origin, approximately 65% of the sperm were attached to OEC within 2h. From 6h of co-culture, the numbers of unattached sperm on ampullary OEC were significantly higher than those on isthmic OEC (p<0.005). From 12h of co-culture, the motility of unattached sperm on isthmic OEC were significantly higher than those on ampullary OEC(p<0.05). The cleavage rate of oocytes inseminated on OEC derived from isthmic segment was also significantly higher than those from ampullary segment (p<0.01). In experiment 2, the numbers of unattached hsp on OEC were significantly higher than those of controls (p<0.01), between 2-24h examination. From 12h of co-culture, the motility of unattached nsp were significantly greater than those of hsp (p<0.01). These results show that bovine OEC derived from the isthmus play more important role(s) for sperm binding, maintaining motility and fertilization in vitro than those from the ampulla, and heparin induced capacitation may change sperm binding ability on OEC in vitro.

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Effects of pH and Carbon Sources on Biohydrogen Production by Co-Culture of Clostridium butyricum and Rhodobacter sphaeroides

  • Lee, Jung-Yeol;Chen, Xue-Jiao;Lee, Eun-Jung;Min, Kyung-Sok
    • Journal of Microbiology and Biotechnology
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    • v.22 no.3
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    • pp.400-406
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    • 2012
  • To improve the hydrogen yield from biological fermentation of organic wastewater, a co-culture system of dark- and photo-fermentation bacteria was investigated. In a pure-culture system of the dark-fermentation bacterium Clostridium butyricum, a pH of 6.25 was found to be optimal, resulting in a hydrogen production rate of 18.7 ml-$H_2/l/h$. On the other hand, the photosynthetic bacterium Rhodobacter sphaeroides could produce the most hydrogen at 1.81mol-$H_2/mol$-glucose at pH 7.0. The maximum specific growth rate of R. sphaeroides was determined to be 2.93 $h^{-1}$ when acetic acid was used as the carbon source, a result that was significantly higher than that obtained using either glucose or a mixture of volatile fatty acids (VFAs). Acetic acid best supported R. sphaeroides cell growth but not hydrogen production. In the co-culture system with glucose, hydrogen could be steadily produced without any lag phase. There were distinguishable inflection points in a plot of accumulated hydrogen over time, resulting from the dynamic production or consumption of VFAs by the interaction between the dark- and photo-fermentation bacteria. Lastly, the hydrogen production rate of a repeated fed-batch run was 15.9 ml-$H_2/l/h$, which was achievable in a sustainable manner.

Lactobacillus acidophilus Contributes to a Healthy Environment for Vaginal Epithelial Cells

  • Pi, Woo-Jin;Ryu, Jae-Sook;Roh, Jae-Sook
    • Parasites, Hosts and Diseases
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    • v.49 no.3
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    • pp.295-298
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    • 2011
  • Lactobacillus species in the female genital tract are thought to act as a barrier to infection. Several studies have demonstrated that lactobacilli can adhere to vaginal epithelial cells. However, little is known about how the adherence of lactobacilli to vaginal epithelial cells affects the acidity, cell viability, or proliferation of the lactobacilli themselves or those of vaginal epithelial cells. Lactobacillus acidophilus was co-cultured with immortalized human vaginal epithelial cells (MS74 cell line), and the growth of L. acidophilus and the acidity of the culture medium were measured. MS74 cell density and viability were also assessed by counting cell numbers and observing the cell attachment state. L. acidophilus showed exponential growth for the first 6 hr until 9 hr, and the pH was maintained close to 4.0-5.0 at 24 hr after culture, consistent with previous studies. The growth curve of L. acidophilus or the pH values were relatively unaffected by co-culture with MS74 cells, confirming that L. acidophilus maintains a low pH in the presence of MS74 cells. This co-culture model could therefore potentially be used to mimic vaginal conditions for future in vitro studies. On the other hand, MS74 cells co-cultured with L. acidophilus more firmly attached to the culture plate, and a higher number of cells were present compared to cells cultured in the absence of L. acidophilus. These results indicate that L. acidophilus increases MS74 cell proliferation and viability, suggesting that lactobacilli may contribute to the healthy environment for vaginal epithelial cells.

Effect of Co-Culture Mouse Fetal Fibroblast Cell on In Vitro Development of Blastomeres Separated from Mouse Preimplantation Embryos (생쥐 태아 Fibroblast 세포와 공동배양이 초기 생쥐배 분할구의 체외 발생능에 미치는 영향)

  • 김진호;정병헌;이훈택;정길생
    • Korean Journal of Animal Reproduction
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    • v.16 no.4
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    • pp.341-346
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    • 1993
  • The development of isolated blastomeres from mammalian preimplantation embryos has been basically studied for the multiplication of embryos from superior animals. Therefore, this study was investigated the effect of co-culture with mouse fetal fibroblast cells(MFFC) on in vitro development of blastomeres from mouse preimplantation embryos. Mature female ICR mice were treated with hormone to induce superovulation and embryos were collected at each 2, 4, and 8-cell stage. Then, after removing zona pellucida with protease, blastomeres were isolated by micropipetting, or reconstituted with different stage blastomere, and incubated for 72 hrs either in T6 or TCM199 or on the monolayer of MFFC, which was prepared with fibroblast cells from 14∼14 day mouse fetus. After incubation, we examined their development rates every day and the nuclei numbers of each blastocyst by Hoechst-33342 staining. In the development rates of blastomeres, there were no significant differences between media but the higher rateswere found in the monolayer of MFFC, regardless of reconsititution. In addition, blastomeres cultured with MFFC had slightly greater number of nuclei than those cultured in single media. Generally, the higher development rates of blastomeres were found from earlier stage embryos than the later ones, regardless of culture conditions. Reconsitituted blastomeres had more nuclei but did not show the higher development rates, compared to the single blastomeres. Taken together, our results suggest that co-culture with MFFC have a beneficial effect on the in vitro development of blastomeres from mouse embryos.

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Preparation of Novel Quaternary Plastomers with Extremely Low Glass Transition Temperature

  • Kim, Jin Hoon;Kim, Jung Soo;Kim, Min Seong;Kim, Ki Bum;Yang, Hong Joo;Ha, Sung Chul;Chang, Young-Wook;Kim, Dong Hyun
    • Elastomers and Composites
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    • v.51 no.3
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    • pp.188-194
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    • 2016
  • In this study, novel quaternary plastomers consisting of ethylene, 1-hexene, high ${\alpha}$-olefin, and divinylbenzene were prepared using Zr metallocene catalyst, borate type cocatalyst, and tri-isobutylaluminium. The molar ratio changes of 1-hexene and high ${\alpha}$-olefin (1-octene, 1-decene, and 1-dodecene) had an effect on the properties of the quaternary plastomers. The structure of the quaternary plastomers was characterized using $^1H$ NMR. Molecular weight properties, crystallinity, and thermal properties of the plastomers were determined by GPC, WAXS, and DMA, respectively. Compared with the terpolymers prepared in our previous study, molecular weight and molecular weight distribution of the quaternary polymers were very similar, whereas glass transition temperature ($T_g$) was very low. Also, hardness and tensile properties of the quaternary plastomers were measured.

ROLE OF FIBROBLASTS IN ORGANOTYPIC CULTURES OF IMMORTALIZED HUMAN ORAL KERATINOCYTES (섬유모세포의 종류에 따른 불멸화된 구강 각화세포의 삼차원적 배양에 관한 연구)

  • Cheong, Jeong-Kwon;Yoon, Kyu-Ho;Kim, Eun-Cheol
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.33 no.3
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    • pp.211-220
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    • 2007
  • Objective: In organotypic culture of immortalized human oral keratinocytes (IHOK), the change of the growth and differentiation was investigated according to the fibroblast type and the involvement of mitogen-activated protein (MAP) kinase. Materials & Methods: IHOK was cultured three dimensionally with gingival fibroblast (GF), dermal fibroblast (DF) and immortalized gingival fibroblast (IGF). We characterized biologic properties of three dimensionally reconstructed IHOK by histological, immunohistochemical, and Western blot analysis. We also investigated whether MAP kinase pathway was involved in epithelial-mesenchymal interaction by Western blot analysis. Results: The best condition of three dimensionally cultured IHOK was the dermal equivalent consisting of type I collagen and IGF. IGF increased the expression of more proliferating cell nuclear antigen (PCNA), involucrin than GF and DF in response to co-culture with IHOK. Extracellularly regulated kinase (ERK) pathway was activated in organotypic co-culture with IGF. Conclusion: The organotypic co-culture of IHOK with dermal equivalent consisting of type I collagen and IGF resulted in excellent morphologic and immunohistochemical characteristics and involved ERK pathway. The epithelial-mesenchymal interaction was activated according to the fibroblast type.