• Asian-Australasian Journal of Animal Sciences
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• v.28 no.8
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• pp.1171-1177
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• 2015

The objective of this study was to establish the optimum protocol for the isolation and culture of porcine muscle satellite cells. Mononuclear muscle satellite cells are a kind of adult stem cell, which is located between the basal lamina and sarcolemma of muscle fibers and is the primary source of myogenic precursor cells in postnatal muscle. Muscle satellite cells are a useful model to investigate the mechanisms of muscle growth and development. Although the isolation and culture protocols of muscle satellite cells in some species (e.g. mouse) have been established successfully, the culture system for porcine muscle satellite cells is very limited. In this study, we optimized the isolation procedure of porcine muscle satellite cells and elaborated the isolation and culture process in detail. Furthermore, we characterized the porcine muscle satellite cells using the immunofluorecence. Our study provides a reference for the isolation of porcine muscle satellite cells and will be useful for studying the molecular mechanisms in these cells.

• The Korean Journal of Physiology and Pharmacology
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• v.24 no.6
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• pp.493-502
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• 2020

Apigenin, a naturally occurring flavonoid, is known to exhibit significant anticancer activity. This study was designed to determine the effects of apigenin on two malignant mesothelioma cell lines, MSTO-211H and H2452, and to explore the underlying mechanism(s). Apigenin significantly inhibited cell viability with a concomitant increase in intracellular reactive oxygen species (ROS) and caused the loss of mitochondrial membrane potential (ΔΨm), and ATP depletion, resulting in apoptosis and necroptosis in monolayer cell culture. Apigenin upregulated DNA damage response proteins, including the DNA double strand break marker phospho (p)-histone H2A.X. and caused a transition delay at the G₂/M phase of cell cycle. Western blot analysis showed that apigenin treatment upregulated protein levels of cleaved caspase-3, cleaved PARP, p-MLKL, and p-RIP3 along with an increased Bax/Bcl-2 ratio. ATP supplementation restored cell viability and levels of DNA damage-, apoptosisand necroptosis-related proteins that apigenin caused. In addition, N-acetylcysteine reduced ROS production and improved ΔΨm loss and cell death that were caused by apigenin. In a 3D spheroid culture model, ROS-dependent necroptosis was found to be a mechanism involved in the anti-cancer activity of apigenin against malignant mesothelioma cells. Taken together, our findings suggest that apigenin can induce ROS-dependent necroptotic cell death due to ATP depletion through mitochondrial dysfunction. This study provides us a possible mechanism underlying why apigenin could be used as a therapeutic candidate for treating malignant mesothelioma.

• Journal of Periodontal and Implant Science
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• v.49 no.3
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• pp.193-204
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• 2019

Purpose: The reaction of cells to a titanium implant depends on the surface characteristics of the implant which are affected by decontamination. The aim of this study was to evaluate the cytocompatibility of titanium disks treated with various decontamination methods, using salivary bacterial contamination with dental pellicle formation as an in vitro model. Methods: Sand-blasted and acid-etched (SA) titanium disks were used. Three control groups (pristine SA disks [SA group]; salivary pellicle-coated SA disks [pellicle group]; and biofilm-coated, untreated SA disks [NT group]) were not subjected to any decontamination treatments. Decontamination of the biofilm-coated disks was performed by 14 methods, including ultrasonic instruments, rotating instruments, an air-powder abrasive system, a laser, and chemical agents. MG63 cells were cultured in the presence of the treated disks. Cell proliferation assays were performed on days 2 and 5 of cell culture, and cell morphology was analyzed by immunofluorescence and scanning electron microscopy (SEM). A vascular endothelial growth factor (VEGF) assay was performed on day 5 of culture. Results: The cell proliferation assay revealed that all decontaminated disks, except for the 2 groups treated using a plastic tip, showed significantly less cell proliferation than the SA group. The immunofluorescence and SEM analyses revealed that most groups showed comparable cell density, with the exception of the NT group, in which the cell density was lower and bacterial residue was observed. Furthermore, the cells grown with tetracycline-treated titanium disks showed significantly lower VEGF production than those in the SA group. Conclusions: None of the decontamination methods resulted in cytocompatibility similar to that of pristine SA titanium. However, many methods caused improvement in the biocompatibility of the titanium disks in comparison with the biofilm-coated, untreated titanium disks. This suggests that decontamination is indispensable for the treatment of peri-implantitis, even if the original biocompatibility cannot be restored.

• Korean Journal of Food Science and Technology
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• v.36 no.2
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• pp.349-354
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• 2004

Predictive growth model of Vibrio parahaemolyticus in modified surimi-based imitation crab broth was investigated. Growth curves of V. parahaemolyticus were obtained by measuring cell concentration in culture broth under different conditions ($Initial\;cell\;level,\;1{\times}10^{2},\;1{\times}10^{3},\;and\;1{\times}10^{4}\;colony\;forming\;unit\;(CFU)/mL$; temperature, 15, 25 37, and $40^{\circ}C$; pH 6, 7, and 8) and applying them to Gompertz model. Microbial growth indicators, maximum specific growth rate (k), lag time (LT), and generation time (GT), were calculated from Gompertz model. Maximum specific growth rate (k) of V. parahaemolyticus increased with increasing temperature, reaching maximum rate at $37^{\circ}C$. LT and GT were also the shortest at $37^{\circ}C$. pH and initial cell number did not influence k, LT, and GT values significantly (p>0.05). Polynomial model, $k=a{\cdot}\exp(-0.5{\cdot}((T-T_{max}/b)^{2}+((pH-pH_{max)/c^{2}))$, and square root model, ${\sqrt{k}\;0.06(T-9.55)[1-\exp(0.07(T-49.98))]$, were developed to express combination effects of temperature and pH under each initial cell number using Gauss-Newton Algorism of Sigma plot 7.0 (SPSS Inc.). Relative coefficients between experimental k and k Predicted by polynomial model were 0.966, 0.979, and 0.965, respectively, at initial cell numbers of $1{\times}10^{2},\;1{\times}10^{3},\;and\;1{\times}10^{4}CFU/mL$, while that between experimental k and k Predicted by square root model was 0.977. Results revealed growth of V. parahaemolyticus was mainly affected by temperature, and square root model showing effect of temperature was more credible than polynomial model for prediction of V. parahaemolyticus growth.

• Journal of Society of Preventive Korean Medicine
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• v.7 no.1
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• pp.55-66
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• 2003

Osteoporosis is characterized by bone loss and mobidity with osteoporotic fracture. This study was performed to evaluate the effect of on the bone mass and its related factors in estrogen-deficient animal model. The model rats of osteoporsis showed a significant decrease in bone density, bone ash density, calcium content of femur bone. At the 14th day after ovariectomy-surgery, rats were administered with DBE, extract of Dioscorea batatas, per orally, and continued for 10 weeks. And osteoporosis related parameters were determined to investigate the effect of DBE. Osteoporetic rats showed lower serum estrogen level, higher body weight than normal rats, and showed atrophy of uterine horns. DBE showed inhibitory effect on bone loss in osteoporetic condition, and reduced the increase of ALP activity and osteocalcin level in serum, and reduced the increase of OH-proline level in urine. But, DBE had no effect on cell proliferation and ALP activity in rat calvarial cell culture.

• Journal of Microbiology and Biotechnology
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• v.12 no.5
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• pp.793-800
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• 2002

The mycolic acids and fatty acids of mycolic acid- containing bacteria in various types of fluids were analyzed using capillary gas chromatography and mass spectrometry. As model strains, Brevibacterium and Coryebacterium species, which have corynomycolic acids ill the range of $C_{32}C_{36}$ in the whole cell, were investigated. Optimized solvents extraction procedures for the mycolic acids and fatty acids from the culture fluids were: chloroform/methanol (1:2, v/v) as the first extraction solvents fur 4 h; and chlorofunuwater (1:1, v/v) as the second extraction solvents far 1 h. These conditions gave above 95% recovery yields fur mycolic acids from the culture fluids. The mycolic acid profile for the whole cells and the culture fluids were similar fur all the media tested. Thus, the procedure described here could be applied for the identification of mycolic acid-containing bacteria in fermentation broth or liquid from of foods.

• Journal of Microbiology and Biotechnology
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• v.10 no.3
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• pp.367-374
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• 2000

The composition of dissolved gases and nutrients in a liquid medium were determined for establishment of the optimum conditions for in vitro culture of Helicobacter pylori. A microaerobic condition facored by the organism was prepared by adjusting the partial pressure of the gas, agitation speed, and viscosity of the medium. The gaseous concentrations were controlled by utilizing CampyPak Plus that reduced oxygen while augmenting carbon dioxide. Agitation of the broth facilitated the oxygen transfer to the cells, yet inhibited the growth at high rates. An increase of viscosity in the medium repressed the culture although this variable was relatively insignificant. The chemical constituents of the liquid broth were examined to establish an economic model for H. pylori cultivation. The microbe required a neutral pH for optimum growth, and yet was also able to proliferate in an acidic condition, presumably by releasing the acidity-modulating enzyme, urease. Cyclodextrin and casamino acid were investigated as growth enhancers in place of serum, while yeast extract unexpectedly inhibited the cells. A low concentration of glucose, the unique carbon source for the organism, increased the cell density, yet high concentrations resulted in an adverse effect. Under optimally dissolved gas conditions, the cell concentration in brucella broth supplemented with serum substitutes and glucose reached $1.6{\times}10^8$ viable cells/ml which was approximately 50% higher than that obtained in the liquid medium added with only cyclodextrin or serum.

• Food Science and Preservation
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• v.16 no.2
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• pp.253-258
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• 2009

This study investigated the effects of mycelium and culture supernatant of Cordyceps ochraceostromat(Co) on air way hyper-responsiveness, pulmonary immune cell infiltration, and Th2 cytokine expression in animal models of atopy and asthma. After ConA(+/-) activation of mouse primary spleen cells, decreased IL-4 and IL-13 cytokine production were seen in the presence of Co mycelium extracts and culture supernatant. The asthma model involved mice sensitized to ovalbumin by i.p. injection treatment; Co mycelium extract was also injected. The atopy model was the dinitrofenylbenzene-treated mouse ear. Ear thicken ing induced by DNFB was decreased by Co mycelium extract, and the extract also inhibited lung cell infiltration in ovalbumin-induced asthmatic mice. The results thus indicated that the Co mycelial extract reduced the undesirable immune responses seen in asthma and atopy.

• Archives of Pharmacal Research
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• v.29 no.5
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• pp.424-429
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• 2006

In the present study, a human mammary epithelial cell (HMEC) culture model was developed to evaluate the potential involvement of carrier-mediated transport systems in drug transfer into milk. Trypsin-resistant HMECs were seeded on $Matrigel^{circledR}-coated$ filters to develop monolayers of functionally differentiated HMEC. Expression of the specific function of HMEC monolayers was dependent of the number of trypsin treatments. Among the monolayers with different numbers of treatment (treated 1 to 3 times), the monolayer treated 3 times (3-t-HMEC monolayer) showed the highest maximal transepithelial resistance and expression of $\beta-casein$ mRNA as an index of differentiation. Transport of tetraethylammonium (TEA) across the 3-t-HMEC monolayer in the basolateral-to-apical direction was significantly higher than that in the apical-to-basolateral direction (p<0.05), whereas such directionality was not observed for p-aminohippurate, suggesting the existence of organic cation transporters, but not organic anion transporters. In fact, expression of mRNAs of human organic cation transporter (OCT) 1 and 3 were detected in the 3-t-HMEC monolayer. These results indicate that the 3-t-HMEC monolayer is potentially useful for the evaluation of carrier-mediated secretion of drugs including organic cations into human milk.

• Reproductive and Developmental Biology
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• v.35 no.1
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• pp.9-15
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• 2011

The functional cardiovascular system is comprised of distinct mesoderm-derived lineages including endothelial cells, vascular smooth muscle cells and other mesenchymal cells. Recent studies in the human embryonic stem cell differentiation model have provided evidence indicating that these cell lineages are developed from the common progenitors such as hemangioblasts and cardiovascular progenitor cells. Also, the studies have suggested that these progenitors have a common primordial progenitor, which expresses KDR (human Flk-1, also known as VEGFR2, CD309). We demonstrate here that sustained activation of BMP4 (bone morphogenetic protein 4) in hESC line, CHA15 hESC results in $KDR^+$ mesoderm specific differentiation. To determine whether the $KDR^+$ population derived from hESCs enhances potential to differentiate along multipotential mesodermal lineages than undifferentiated hESCs, we analyzed the development of the mesodermal cell types in human embryonic stem cell differentiation cultures. In embryoid body (EB) differentiation culture conditions, we identified an increased expression of $KDR^+$ population from BMP4-stimulated hESC-derived EBs. After induction with additional growth factors, the $KDR^+$ population sorted from hESCs-derived EBs displays mesenchymal, endothelial and vascular smooth muscle potential in matrix-coated monolayer culture systems. The populations plated in monolayer cultures expressed increased levels of related markers and exhibit a stable/homologous phenotype in culture terms. In conclusion, we demonstrate that the $KDR^+$ population is stably isolated from CHA15 hESC-derived EBs using BMP4 and growth factors, and sorted $KDR^+$ population can be utilized to generate multipotential mesodermal progenitors in vitro, which can be further differentiated into cardiovascular specific cells.