• Title/Summary/Keyword: optimization of culture conditions

Search Result 313, Processing Time 0.033 seconds

Optimization of Culture Conditions for Maintaining Pig Muscle Stem Cells In Vitro

  • Choi, Kwang-Hwan;Yoon, Ji Won;Kim, Minsu;Jeong, Jinsol;Ryu, Minkyung;Park, Sungkwon;Jo, Cheorun;Lee, Chang-Kyu
    • Food Science of Animal Resources
    • /
    • v.40 no.4
    • /
    • pp.659-667
    • /
    • 2020
  • Muscle stem cells isolated from domestic animals, including cows and pigs, were recently spotlighted as candidates for the production of alternative protein resources, so-called cultured meat or lab-grown meat. In the present study, we aimed to optimize the in vitro culture conditions for the long-term expansion of pig muscle stem cells via the screening of various signaling molecules. Pig muscle stem cells were collected from the biceps femoris muscles of 3-d-old crossbred pigs (Landrace×Yorkshire×Duroc, LYD) and cultured in minimum essential medium-based growth media. However, the pig muscle stem cells gradually lost their proliferation ability and featured morphologies during the long-term culture over two weeks. To find suitable in vitro culture conditions for an extended period, skeletal muscle growth medium-2, including epidermal growth factor (EGF), dexamethasone, and a p38 inhibitor (SB203580), was used to support the stemness of the pig muscle stem cells. Interestingly, pig muscle stem cells were stably maintained in a long-term culture without loss of the expression of myogenic marker genes as determined by PCR analysis. Immunostaining analysis showed that the stem cells were capable of myogenic differentiation after multiple passaging. Therefore, we found that basal culture conditions containing EGF, dexamethasone, and a p38 inhibitor were suitable for maintaining pig muscle stem cells during expanded culture in vitro. This culture method may be applied for the production of cultured meat and further basic research on muscle development in the pig.

Optimization of Culture Conditions and Analysis of Plasmid Stability of a Transformant Bacillus subtilis for Cytidine Deaminase Production

  • Kim, Soo-Hyun;Song, Bang-Ho;Lee, Yong-Hyun
    • Journal of Microbiology and Biotechnology
    • /
    • v.1 no.2
    • /
    • pp.116-120
    • /
    • 1991
  • The transformant Bacillus subtilis ED213 carrying the pSO100 which cloned the cdd gene encoding cytidine deaminase (cytidine /2'-deoxycytidine aminohydrolase, EC 3.5.4.5, CDase) originated from wild type B. subtilis was cultivated in Spizizen minimal medium (SMM). To overcome poor expression of the cdd gene in SMM medium, the medium compositions and growth conditions were optimized. The optimized medium compositions and growth conditions were cytidine concentration of 80 mg/l, glycerol of 25 g/l, and $(NH_4)_2SO_4$ of 10 g/l, along with $37^{\circ}C$ and pH 7.0. The intracellular CDase production was increased 3 times from 1,000 unit/ml to 3,200 unit/ml, and extracellular CDase also increased from nearly undetectable amounts to 1,500 unit/ml. The cytidine concentration was signified as the most critical compositional factor for overproduction of CDase by increasing the cell density mainly in culture broth. The plasmids were more stable in cells that were grown in original SMM medium with stability of 90% compared to those grown in optimized SMM medium with stability of 80% after 48 hours cultivation. The most active amplification of plasmid was occurred in the logarithmic phase, which showed a value around four times higher than the initial copy number. In the exponential phase, the CDase production was closely related to the plasmid copy number along with the cell density. However, it was not accorded with cell density at the stationary phase.

  • PDF

The Improvement of Cephalosporin C Production by Fed-batch Culture of Cephalosporium acremonium M25 Using Rice Oil

  • Kim Jin Hee;Lim Jung Soo;Kim Seung Wook
    • Biotechnology and Bioprocess Engineering:BBE
    • /
    • v.9 no.6
    • /
    • pp.459-464
    • /
    • 2004
  • The objective of this study is to improve cephalosporin C (CPC) production byoptimization of medium and culture conditions. A statistical method was introduced to optimize the main culture medium. The main medium for CPC production was optimized using a statistical method. Glucose and corn steep liquor (CSL) were found to be the most effective factors for CPC production. Glucose and CSL were optimized to 2.84 and $6.68\%$, respectively. CPC produc­tion was improved $50\%$ by feeding of $5\%$ rice oil at day 3rd and 5th day during the shake flask culture of C acremonium M25. The effect of agitation speeds on CPC production in a 2.5-L bio­reactor was also investigated with fed-batch mode. The maximum cell mass (54.5 g/L) was obtained at 600 rpm. However, the maximum CPC production (0.98 g/L) was obtained at 500 rpm. At this condition, the maximum CPC production was improved about $132\%$ compared to the re­sult with batch flask culture.

Isolation and identification of Exo-Inulinase Producing Bacterium and Optimization of the Enzyme Production (Exo-inulinase 생산 균주의 분리ㆍ동정 및 효소 생산의 최적화)

  • 김병우;이경희
    • Journal of Life Science
    • /
    • v.9 no.1
    • /
    • pp.22-28
    • /
    • 1999
  • A bacterium producing exo-inulinase was isolated from soil and identified Pseudomonas sp. and named as Pseudomonas sp. NO5. The optimal culture conditions for the efficient production of exo-inulinase from Pseudomonas sp. NO5 were obtained by cultivating with the medium 1$\%$ sucrose, 0.5$\%$ yeast extract, 0.5$\%$ $(NH_4)_2$$HPO_4$, 0.05$\%$ $MgSO_4$$7H_2$0, 0.001$\%$ and $FeSO_4$$7H_2$0 at $37^{\circ}C$ in initial pH 7.0 for 20 hours. The enzyme was induced maximally in the presence of sucrose or inulin at early stationary phase about 20 hour after cultivation.

  • PDF

Development of Eco-friendly Reduction Process for Indigo Dyeing : Using Hansenula misumaiensis Strain (인디고 염색을 위한 친환경 환원공정 개발: 한세눌라 균주의 이용)

  • Shin, Younsook;Son, Kyunghee;Yoo, Dong Il
    • Textile Coloration and Finishing
    • /
    • v.26 no.3
    • /
    • pp.237-241
    • /
    • 2014
  • The aim of this study is to develop an eco-friendly reduction process of indigo as an alternative choice. Hansenula misumaiensis was used and their reducing activity toward synthetic indigo as well as natural indigo was estimated by dyeing test in terms of indigo dye uptake. The changes in K/S value and pH were monitored on the time-based measurements. Also, reduction duration was evaluated. On the basis of the results described in this study, it was confirmed that Hansenula misumaiensis reduced indigo. Reducing power of Hansenula misumaiensis reached to maximum in two days. It can be possible to develop eco-friendly process of indigo reduction using Hansenula misumaiensis by the optimization of strain culture conditions and the optimization of reduction conditions.

Isolation of Bacillus sp. Producing Multi-enzyme and Optimization of Medium Conditions for Its Production Using Feedstuffs for Probiotics (Probiotics용 복합효소 분비 Bacillus sp.의 분리 및 원료사료를 이용한 균주 생산을 위한 배지 조건의 최적화)

  • 양시용;송민동;김언현;김창원
    • Microbiology and Biotechnology Letters
    • /
    • v.29 no.2
    • /
    • pp.110-114
    • /
    • 2001
  • Isolation of BacilLus sp. producing multi-enzyme and optimization of medium conditions for its production using feedstuffs for probiotics were carried out in this study. A bacterium isolated from natural resources, namely Bacillus subtilis 4-3, has multi-enzyme activity (phytase. cellulase, xylanasc, protease, and amylase. In the culture of B. subtilis 4-3 using soybean meal and rice bran. relatively low phytate degradation was noted using whereas high phytate degradability was observed with wheat bran (80.63%). The optimal composition of medium using feedstuffs was 1.0% (w/v) soybean meal and 2% (w/v) molasses to yield high cell growth.

  • PDF

Optimization of the in vitro fertilization system in pigs

  • Song-Hee Lee;Xiang-Shun Cui
    • Journal of Animal Reproduction and Biotechnology
    • /
    • v.38 no.2
    • /
    • pp.70-76
    • /
    • 2023
  • Background: Despite considerable technological advancements, polyspermy remains a significant challenge in in vitro fertilization (IVF) procedures in pigs, disrupting normal embryonic development. Here, we aimed to determine whether optimal fertilization conditions reduce the polyspermy incidence in pigs. Methods: In vitro-matured oocytes were co-incubated with sperm according to a modified two-step culture system. Results: In the first experiment, oocytes were briefly co-incubated with sperm, washed in IVF medium, and then moved to fresh IVF medium for 5 or 6 h. Although the 6 h sperm-free cultured group had a higher penetration rate than the 5 h cultured group, the polyspermy rate significantly increased in the 6 h sperm-free cultured group. The gamete co-incubation period was either 20 or 40 min. The 40 min cultured group had a higher rate of blastocyst formation and number of total cells in blastocysts than the 20 min cultured group. In experiment 2, oocytes were inseminated with sperm separated by Pecroll treatment. Percoll treatment increased the rate of oocyte penetration and blastocyst formation compared to the control. In experiment 3, fertilized oocytes were cultured in 25 µL microdroplets (10 gametes/drop) or 500 µL (100 gametes/well) of culture medium in 4-well plates. The large volume of medium significantly reduced the number of dead oocytes and increased the rate of blastocyst formation compared to the small volume. Conclusions: Collectively, these results demonstrate that various fertilization conditions, including modified co-culture period, active sperm separation, and culture medium volume, enhance fertilization efficiency and subsequent embryonic development by decreasing polyspermy occurrence.

The Optimization of Muffin with Yam Powder Using Response Surface Methodology (마분말 첨가 머핀 제조조건 최적화)

  • Joo, Na-Mi;Lee, Sun-Mee;Jeong, Hee-Sun;Park, Sang-Hyun;Jung, Ah-Ram;Ryu, Seung-Yeon;Lee, Ji-Hee;Jung, Hyeon-A
    • Journal of the Korean Society of Food Culture
    • /
    • v.23 no.2
    • /
    • pp.243-251
    • /
    • 2008
  • This purpose of this study was to develop a functional muffin by adding yam powder in the shape of a muffin as a partial surrogate for wheat flour. The yam has been found to be effective for liver and kidney function, as well as the digestion of protein, since it produces glucuronic acid in the body. Therefore, the purpose of this study was to determine the optimal mixing conditions of yam muffins by adjusting the amounts yam powder, butter, and sugar. The mixing conditions for the yam muffins included 3 categories: yam powder $(X_1)$, sugar $(X_2)$, and butter $(X_3)$ by Central Composite Design (CCD) which was optimized by Response Surface Methodology (RSM). The effects of the three variable additions on muffin quality were examined via physical and chemical experiments, such as the analysis of texture (hardness, cohesiveness, springiness, gumminess), coloration (lightness, redness, yellowness), and height. Lastly, we performed a sensory test, which revealed significant findings for gumminess, color, appearance, flavor, softness (p<0.05), redness, and overall quality (p<0.01). Consequently, the optimal mixing rate which best satisfied the sensory items were 34.35g of yam powder, 80.15 g of sugar, and 80.55 g of butter.

Isolation of Pigment Overproducing Mutant from Monascus purpureus and Optimization of Pigment Production (Monascus purpureus로부터 다량의 색소생성 변이주의 분리 및 색소생성의 최적조건)

  • Park Chi Duck;Jung Hyuck Jun;Yu Tae Shick
    • Korean Journal of Microbiology
    • /
    • v.41 no.2
    • /
    • pp.130-134
    • /
    • 2005
  • Isolation of a pigment overproducing mutant, P-57, by ultraviolet irradiation of Monascus purpureus KCCM 60016 and investigation of the optimal conditions for pigment production of the mutant were carried out. P-57 mutant produced pigment on solid state culture. Unpolished rice was the best cereal source for pigment production among eight kinds of tested cereal sources for the solid culture of the mutant. The optimal culture condition for pigment production were obtained from the cultivated at $30^{\circ}C,\;90\%$ humidity for 30 days. The P-57 mutant strain showed the best pigment productivity of 160.0 unit at red pigment, 193.6 unit at orange pigment, and 141.6 unit at yellow pigment on solid state culture under optimal condition.

Optimization of Monochamus alternatus media and culture period for cordycepin production in Cordyceps militaris culture using solid-state fermentation

  • Ha, Si Young;Jung, Ji Young;Yang, Jae-Kyung
    • Journal of Mushroom
    • /
    • v.19 no.3
    • /
    • pp.126-133
    • /
    • 2021
  • In this study, we investigated the effect of solid culture medium on the production of cordycepin in Cordyceps militaris. The regression equation was expressed as follows: Y1 = 755.3-58.6625X1+4.79432E-14X2-46.6625X3-5.66269E-14X1X2-0.025X1X3+1.62475E-14X2X3-160.6625X12+0.0125X22-206.9625X32, where, Y represents the value of cordycepin content (㎍/g), X1 corresponds to the weight of M. alternatus in solid culture medium (g/bottle), X2 to the water content of the solid culture medium (%), and X3 to the culture period (day). The solid culture medium was optimized using the response surface methodology, and the optimal medium composition was as follows: the weight of M. alternatus in solid culture medium and water content were 16.2% and 100.7% (20.14 mL water/20 g solid culture medium), respectively, with a culture period of 39 days. Under these conditions, the cordycepin content of the fruiting bodies reached 150.0 ㎍/g (actual value). The supplementation of M. alternatus in solid culture for improved cordycepin content of C. militaris seems to be a promising alternative to wild and solid cultivation.