• 한국축산식품학회지
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• 제43권6호
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• pp.1017-1030
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• 2023

Fetal bovine serum (FBS), which contains various nutrients, comprises 20% of the growth medium for cell-cultivated meat. However, ethical, cost, and scientific issues, necesitates identification of alternatives. In this study, we investigated commercially manufactured serum-free media capable of culturing Hanwoo satellite cells (HWSCs) to identify constituent proliferation enhancing factors. Six different serum-free media were selected, and the HWSC proliferation rates in these serum-free media were compared with that of control medium supplemented with 20% FBS. Among the six media, cell proliferation rates were higher only in StemFlex^TM Medium (SF) and Mesenchymal Stem Cell Growth Medium DXF (MS) than in the control medium. SF and MS contain high fibroblast growth factor 2 (FGF2) concentrations, and we found upregulated FGF2 protein expression in cells cultured in SF or MS. Activation of the fibroblast growth factor receptor 1 (FGFR1)-mediated signaling pathway and stimulation of muscle satellite cell proliferation-related factors were confirmed by the presence of related biomarkers (FGFR1, FRS2, Raf1, ERK, p38, Pax7, and MyoD) as indicated by quantitative polymerase chain reaction, western blotting, and immunocytochemistry. Moreover, PD173074, an FGFR1 inhibitor suppressed cell proliferation in SF and MS and downregulated related biomarkers (FGFR1, FRS2, Raf1, and ERK). The promotion of cell proliferation in SF and MS was therefore attributed to FGF2, which indicates that FGFR1 activation in muscle satellite cells may be a target for improving the efficiency of cell-cultivated meat production.

• 한국가축번식학회지
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• 제20권1호
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• pp.77-84
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• 1996

본 연구는 배양액의 삼투압과 비타민이 체외 배발생에 미치는 영향을 조사하기 위하여 소의 미성숙 난포란을 체외성숙 및 수정 후 무단백 한정배양액에서 체외배양하였다. 0.35mM phosphate와 19종 아미노산이 포함된 무단백 한정배양액의 삼투압을 NaCl농도에 의하여 달리하였을 때 소 수정란의 발생능력은 265mOsm과 290mOsm에서 유의적으로 증가하였다(p<0.05). 수정 후 120시간에 5.56mM포도당이 포함된 새로운 배양액에서 배발생을 계속시켰을 때 상실배는 290mOsm과 315mOsm에서 유의적으로 증가하였으며, 배반포는 290mOsm배양액에서 유의적으로 증가하였다. Glutamine과 아미노산이 첨가된 배양액에서 비타민은 소수정란의 발생에 영향을 미치지 않았다.

• 대한생식의학회:학술대회논문집
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• 대한불임학회 2004년도 제47차 추계학술대회
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• pp.54.1-54.1
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• 2004

• Biotechnology and Bioprocess Engineering:BBE
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• 제11권4호
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• pp.319-324
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• 2006

Although the sera used in animal cell culture media provide the macromolecules, nutrients, hormones, and growth factors necessary to support cell growth, it could also be an obstacle to the production of recombinant proteins in animal cell culture systems used in many sectors of the biotechnology industry. For this reason, many research groups, including our laboratory, have been trying to develop serum-free media (SFM) or serum-supplemented media (SSM) for special or multi-purpose cell lines. The Chinese hamster ovary (CHO) cell, for example, is frequently used to produce proteins and is especially valuable in the large-scale production of pharmaceutically important proteins, yet information about its genome is lacking. Also, SFMs have only been evaluated by comparing growth patterns for cells grown in SFMs with those grown in SSM or by measuring the titer of the target protein obtained from cells grown in each type of medium. These are not reliable methods of obtaining the type of information needed to determine whether an SFM should be replaced with an SSM. We carried out a cDNA microarray analysis to evaluate MED-3, an SFM developed in our laboratory, as a CHO culture medium When CHO cells were cultured in MED-3 instead of an SSM, several genes associated with cell growth were down-regulated, although this change diminished over time. We found that the insulin-like growth factor (IGF) gene was representative of the proteins that were down-regulated in cells cultured in MED-3. When several key supplements - including insulin, transferrin, ethanolamine, and selenium - were removed from MED-3, the IGF expression was consistently down- regulated and cell growth decreased proportionately. Based on these results, we concluded that when an SFM is used as a culture medium, it is important to supplement it with substances that can help the cells maintain a high level of IGF expression. The data presented in this study, therefore, might provide useful information for the design and development of SFM or SSM, as well as for the design of genome-based studies of CHO cells to determine how they can be used optimally for protein production in pharmaceutical and biomedical research.

• Journal of Microbiology and Biotechnology
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• 제21권8호
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• pp.791-795
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• 2011

Sea tangle, a kind of brown seaweed, was fermented with Lactobacillus brevis BJ-20. The gamma-aminobutyric acid (GABA) content in fermented sea tangle (FST) was 5.56% (w/w) and GABA in total free amino acid of FST was 49.5%. The effect of FST on the enzyme activities and mRNA protein expression of alcohol dehydrogenase (ADH) and acetaldehyde dehydrogenase (ALDH) involved in alcohol metabolism in Saccharomyces cerevisiae was investigated. Yeast was cultured in YPD medium supplemented with different concentrations of FST powder [0, 0.4, 0.8, and 1.0% (w/v)] for 18 h. FST had no cytotoxic effect on the yeast growth. The highest activities and protein expressions of ADH and ALDH from the cell-free extracts of S. cerevisiae were evident with the 0.4% and 0.8% (w/v) FST-supplemented concentrations, respectively. The highest concentrations of GABA as well as minerals (Zn, Ca, and Mg) were found in the cell-free extracts of S. cerevisiae cultured in medium supplemented with 0.4% (w/v) FST. The levels of GABA, Zn, Ca, and Mg in S. cerevisiae were strongly correlated with the enzyme activities of ADH and ALDH in yeast. These results indicate that FST can enhance the enzyme activities and protein expression of ADH and ALDH in S. cerevisiae.

• Biotechnology and Bioprocess Engineering:BBE
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• 제8권2호
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• pp.142-146
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• 2003

Three insect cell lines, Sf9, Sf21 and Tn5Bl-4, and four different kinds of serum free media (SFM), Sf 900 II, EX-CELL 420, EX-CELL 405 and Express Five, were used to compare the nutrient consumption, byproduct formation, production of recombinant protein and protease activity in suspension cultures. The Sf 900 II SFM was a ppropriate for the cell growth and protein production of the Sf9 and Sf21 cell lines. When the Tn5Bl-4 cell line was grown in the Express Five SFM, the specific growth rate was 1.6 fold higher than those of either the Sf9 or Sf21 cell lines. The glucose and glutamine consumption rates per cells, were 4 and 2.3 times higher than those of the Sf9 cell line, respectively. The overall yield coefficients of the lactate and ammoniumion were 2.8 and 1.5 times higher compared to those of the Sf9 cell line. respectively. The maximum specific ${\beta}$-galactosidase production rate was 4.5 fold that of the Sf9 cell line, a 3 times higher protease activity per cell.

• Plant Resources
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• 제1권1호
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• pp.1-5
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• 1998

Total iron content and ferritin distribution have been determined in red pepper(Capsicum annuum L.) during development stage under conditions of iron nutritional status from hydroponic culture. Color of the leaves become chlorotic on iron deficient and high concentration. The plant height on each iron concentration had retarding effect at concentration lower than $25\muM$ and greater than 125$25\muM$. In normal green leaves. Total iron content was almost constant with a mean value of $2.5\mumole$ of iron/mg of dry matter, except at 63day, for which it increases slightly to $4\mumole$. Howere, iron content of chlorotic plants grew on iron free medium was not almost detectable. Also in post chlorotic leaves(++Fe), iron content was evidently increase unitl 7days after transfer on liquid medium, but decreased from after 14days. Also, ferritin protein analysed total protein extracts prepared from leaves of different ages using antibodies raised against ferritin protein. Ferritin protein deereased progressively during the first week of germination and was not detectable in vegetative tissues. Ferritin protein in post chlorotic leaves wasevidently strongly cnhanced until 11days after transfer on liquid medium but decreased until the leves became chlorotic.

• Plant Resources
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• 제5권1호
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• pp.29-44
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• 2002

In this study, plant regeneration through in vitro culture from plantlet stems of Yooja (C. junos Sieb.) and trifoliate orange (P. trifoliata Rafin.) was attempted to make mass-production system of virus-free plants having the same genotype with mother plant. In order to investigate physiological change depending on the developmental stage of plant regeneration, the changes of total protein, peroxidase and esterase activity and their isozyme patterns as well were examined in 1/2 MS medium. The results are as follows : 1. The MS medium for the optimal callus induction and shoot formation was utilized. The medium was supplemented either with 2,4-D and Kinetin or with BA and NAA. The optimal concentrations were the combination of 1.0mg/ 2,4-D +0.3mg/ Kinetin and 1.0mg BA +0.3mg NAA in callus induction and shoot formation, respectively. 2. For the plant regeneration from somatic embryos, 1/2 MS medium was used with supplements of growth regulators (free, 1.0mg/ IBA +1.0mg/ BA ,0.5mg/ IBA +0.5mg/ BA). Shooting and rooting were the best in the treatment of 0.5mg/ IBA and 0.5mg/ BA combination. 3. The total protein content has a tendency of increase with the developmental stage of embryo, but it was decreased at the plantlet. Also it was the highest at 8 and 6 weeks stage in C. junos Sieb. and P. trioliata Rafin, respectively. In the SDS-PAGE pattern of protein, C. junos Sieb. showed bands of 29.0 and 40kDa at 10 weeks. The 45,66 and 97.4 kDa bands at 10 weeks of culture were shown in P. trifoliata Rafin. 4. The highest esterase activity was shown at the 6 and 8 weeks of culture in C.junos Sieb. and P. trifoliata Rafin.., respectively. 5. Esterase isozyme patterns were shown difference according to the developmental stage. In C. junos Sieb. a new band was observed at pl 7.7 following 4 weeks culture. On the other hand, new bands in P. trifoliata Rafin. were observed at pl 7.5~6.5 following 4 and 6 weeks culture, respectively.

• 한국식품영양과학회지
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• 제25권3호
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• pp.513-518
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• 1996

In order to investigate the characteristics of alcohol fermentation by Saccharomyces cerevisiae, compositions and free sugar content of dried riped pumpkin powder were determined. The proximate compositions of ripe pumpkin powder were as follows: moisture 11.98%, total sugar 62.18%(dried weight basis 70.64%), protein 13.38%, lipid 0.85%, fiber 7.07% and ash 4.54%. The compositions of free sugar in well ripe pumpkin were: glucese 85.36mg/g, fructose 40.68mg/g, sucrose 68.25mg/g, lactose 18.60mg/g and maltose 3.82mg/g. The optimum conditions for alcohol fermentation by S. cerevisiae were as follows; incubation temperature of 3$0^{\circ}C$, initial pH of 6.0, ripe pumpkin powder concentration of 10% and cells inoculation of 1.3$\times$$10^{6}$ cells/ml liquid medium. Ethanol production under the optimum conditions was 5.95g/100g in liquid medium containing 10% ripe pumpkin powder after 4 days incubation.

• Smart Structures and Systems
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• 제26권6호
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• pp.809-817
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• 2020

In cells, the microtubules are surrounded by viscoelastic medium. Microtubules, though very small in size, perform a vital role in transportation of protein and in maintaining the cell shape. During performing these functions waves propagate and this propagation of waves has been investigated using nonlocal elastic theory. But the effect of surrounding medium was not taken into account. To fill this gap, this study considers the viscoelastic medium along with nonlocal elastic theory. The analytical formulas of the velocity of waves, and the results reveal that the presence of medium reduces the velocity. The axisymmetric and nonaxisymmetric waves are separately discussed. Furthermore, the results are compared with the results gained from the studies of free microtubules. The presence of medium around microtubules results in the increase of the flexural rigidity causing a significant decrease in radial wave velocity as compared to axial and circumferential wave velocities. The effect of viscoelastic medium is more obvious on radial wave velocity, to a lesser extent on torsional wave velocity and least on longitudinal wave velocity.