Cell Culture Solution Research Articles

Colorimetric polymerase chain reaction mediated by pH indicator: Rapid detection of drug-resistant nosocomial bacteria

Polymerase chain reaction (PCR) is the gold standard for molecular diagnoses due to its high sensitivity and accuracy. However, conventional PCR exhibits limitations including extended analysis time and complicated sample treatment. To shorten the analysis time of standard three-step PCR, we employed a two-step PCR allowing for fast amplification of nucleic acid in a chip format. Moreover, to realize a fully-integrated and on-site visual discrimination of opportunistic and multidrug-resistant pathogens, namely, Enterococcus faecium and Acinetobacter baumannii. DNA was extracted using FTA card, and pH-dependent color changes of the PCR amplicons were monitored by using phenolphthalein as a pH indicator. The overall reaction took less than 35 min and exhibited high selectivity and sensitivity confirmed by the limit of detection of approximately 103 CFU/mL for E. faecium. In addition, carbapenem-resistant A. baumannii was successfully detected demonstrating the clinical applicability of the introduced technique using a thermoplastic chip. E. faecium both in bacterial cell culture solution and a contaminated surface were also successfully detected proving the feasibility of the chip-based detection system and paving the way for a facile discrimination of opportunistic pathogens prevalent in hospitals and our environment.

The superiority of innovative spiral-interdigital microelectrode pattern in increasing the sensitivity of tracing synchronization via serum starvation in cellular metabolism

In order to investigate the changes in the properties of the cell culture solution in the effect of cell synchronization via cell starvation (for 12, 24, and 36 h), a new spiral-interdigital pattern of microelectrode as a biosensor has been proposed. Then, to test its superiority, the results of this spiral-interdigital pattern with the results of the commercial pattern have been compared. The cells were selected from breast cancer standard lines (MDA-MB-231). Changes in CV peaks of the secretions were recorded by the spiral-interdigital pattern, in which increasing the interactive surface with homogenous electric paths had been considered by simulation before fabrication. The results of the simulation and experimental procedures showed a meaningful correlation. The occurrence of CV oxidative peaks at about 0.1–0.4 V and reductive peaks at approximately 0 V in the spiral-interdigital biosensor in the starved MDA-MB-231 cell line has been observed. The starvation situation resembles one that does not cause meaningful cell apoptosis or necrosis, and this method is only used to make the cells synchronized. Also, no peak is observed in normal cell growth conditions. In addition, by using the commercial design of the electrodes, no peak is observed in any of the conditions of normal and synchronized growth of the cells. Therefore, it seems that the observed peaks are caused by the agents that are secreted in the cell culture solution in a synchronized situation. Moreover, the design of the new spiral-interdigital electrode can significantly increase the sensitivity of the sensor to receive these peaks due to more space and a uniform electric field.

Curcumin-activated Wnt5a pathway mediates Ca2+ channel opening to affect myoblast differentiation and skeletal muscle regeneration.

Skeletal muscle injury is one of the most common sports injuries; if not properly treated or not effective rehabilitation treatment after injury, it can be transformed into chronic cumulative injury. Curcumin, an herbal ingredient, has been found to promote skeletal muscle injury repair and regeneration. The Wnt5a pathway is related to the expression of myogenic regulatory factors, and Ca2+ promotes the differentiation and fusion process of myoblasts. This study explored the effect and mechanism of curcumin on myoblast differentiation during the repair and regeneration of injured skeletal muscle and its relationship with the Wnt5a pathway and Ca2+ channel. Myogenic differentiation of C2C12 cells was induced with 2% horse serum, and a mouse (male, 10weeks old) model of acute skeletal muscle injury was established using cardiotoxin (20μL). In addition, we constructed a Wnt5a knockdown C2C12 cell model and a Wnt5a knockout mouse model. Besides, curcumin was added to the cell culture solution (80mg/L) and fed to the mice (50mg/kg). Fluorescence microscopy was used to determine the concentration of Ca2+. Western blot and RT-qPCR were used to detect the protein and mRNA levels of Wnt5a, CaN, NFAT2, MyoD, Myf5, Pax7, and Myogenin. The expression levels of MyoD, Myf5, Myogenin, MHC, Desmin, and NFAT2 were detected using immunofluorescence techniques. In addition, MyoD expression was observed using immunohistochemistry, and morphological changes in mouse muscle tissue were observed using HE staining. During myoblast differentiation and muscle regeneration, Wnt5a expression was upregulated (P<0.001) and the Wnt5a signalling pathway was activated. Wnt5a overexpression promoted the expression of MyoD, Myf5, Myogenin, MHC, and Desmin (P<0.05), and conversely, knockdown of Wnt5a inhibited their expression (P<0.001). The Wnt5a pathway mediated the opening of Ca2+ channels, regulated the expression levels of CaN, NFAT2, MyoD, Myf5, Myogenin, MHC, and Desmin (P<0.01) and promoted the differentiation of C2C12 myoblasts and the repair and regeneration of injured skeletal muscle. The expression of Wnt5a, CaN, NFAT2, MyoD, Myogenin, Myf5, and MHC in C2C12 myoblast was significantly increased after curcumin intervention (P<0.05); however, their expression decreased significantly after knocking down Wnt5a on the basis of curcumin intervention (P<0.05). Similarly, in Wnt5a knockout mice, the promotion of muscle regeneration by curcumin was significantly attenuated. Curcumin can activate the Wnt5a signalling pathway and mediate the opening of Ca2+ channels to accelerate the myogenic differentiation of C2C12 cells and the repair and regeneration of injured skeletal muscle.

The Formation of Bixbyite-Type Mn2O3 via Pyocyanin-Dependent Mn(II) Oxidation of Soil-Derived Pseudomonas aeruginosa

Bacterial Mn(II) oxidation is believed to play a dominant role in accelerating the rate of Mn biomineralization in nature. Commonly, bacteria employ two ways involving Mn(II) oxidases and reactive oxygen species to oxidize Mn(II). In this study, a new strategy for bacterial Mn(II) oxidation involving the pyocyanin, a greenish blue phenazine pigment from Pseudomonas aeruginosa, was discovered. To begin with, a bacterial strain L3 was isolated from soils and identified as Pseudomonas aeruginosa, which exhibited the ability of Mn(II) oxidation. Next, the pyocyanin was purified from strain L3 cultures and proven to be involved in Mn(II) oxidation. Particularly, the oxidation of Mn(II) by pyocyanin was dependent on its ambient pH. In comparison with pH of 5 and 7, pyocyanin (the initial value of OD387 was 0.56 at pH of 2) showed a stronger capability of oxidizing Mn(II) at pH of 9, reaching 144.03 μg L−1 of Mn oxides after 108 h of Mn(II) oxidation, while pyocyanin ultimately produced 43.81 μg L−1 at pH of 7 and 3.32 μg L−1 at pH of 5, respectively. Further, strain L3 cultures were fractionated into three parts, i.e., the cell culture solution, fermentation supernatant, and cell suspension, and the Mn(II)-oxidizing activity was found to be distributed in the cell culture solution and fermentation supernatant, as evidenced by the formation of blackish glossy Mn oxides. Specifically, in the first half, the rate of Mn(II) oxidation by the fermentation supernatant was higher than that by the cell culture solution, whereas in the second half, the cell culture solution showed the much higher Mn(II)-oxidizing activity compared to the fermentation supernatant. Last but not least, the collective results from mineral characterization demonstrated that, the Mn oxides produced by P. aeruginosa strain L3, either by the cell culture solution or by the fermentation supernatant, were bixbyite-type Mn2O3 with poor crystallinity.

Integrated Sandwich-Paper 3D Cell Sensing Device to In Situ Wirelessly Monitor H2O2 Released from Living Cells.

Point-of-care testing (POCT) has attracted great interest because of its prominent advantages of rapidness, precision, portability, and real-time monitoring, thus becoming a powerful biomedical device in early clinical diagnosis and convenient medical treatments. However, its complicated manufacturing process and high expense severely impede mass production and broad applications. Herein, an innovative but inexpensive integrated sandwich-paper three-dimensional (3D) cell sensing device is fabricated to in situ wirelessly detect H2O2 released from living cells. The paper-based electrochemical sensing device was constructed by a sealed sandwiched bottom plastic film/fiber paper/top hole-centered plastic film that was printed with patterned electrodes. A new (Fe, Mn)3(PO4)2/N-doped carbon nanorod was developed and immobilized on the sensing carbon electrode while cell culture solution filled the exposed fiber paper, allowing living cells to grow on the fiber paper surrounding the electrode. Due to the significantly shortening diffusion distance to access the sensing sites by such a unique device and a rationally tuned ratio of Fe2+/Mn2+, the device exhibits a fast response time (0.2 s), a low detection limit (0.4 μM), and a wide detection range (2-3200 μM). This work offers great promise for a low-cost and highly sensitive POCT device for practical clinic diagnosis and broad POCT biomedical applications.

Plasma-Activated Media Produced by a Microwave-Excited Atmospheric Pressure Plasma Jet Is Effective against Cisplatin-Resistant Human Bladder Cancer Cells In Vitro.

Media exposed to atmospheric pressure plasma (APP) produce reactive oxygen and nitrogen species (RONS), with hydrogen peroxide (H2O2), nitrite (NO2-), and nitrate (NO3-) being among the most detected species due to their relatively long lifetime. In this study, a standardized microwave-excited (ME) APP jet (APPJ) source was employed to produce gaseous RONS to treat liquid samples. The source was a commercially available plasma jet, which generated argon plasma utilizing a coaxial transmission line resonator at the operating frequency of 2.45 GHz. An ultraviolet-visible spectrophotometer was used to measure the concentrations of H2O2 and NO3- in plasma-activated media (PAM). Three different types of media (deionized water, Hank's balanced salt solution, and cell culture solution Dulbecco's modified eagles medium [DMEM]) were utilized as liquid samples. Among these media, the plasma-treated DMEM was observed to have the highest levels of H2O2 and NO3-. Subsequently, the feasibility of using argon ME-APPJ-activated DMEM (PAM) as an adjuvant to enhance the therapeutic effects of cisplatin on human bladder cancer cells (T-24) was investigated. Various cancer cell lines, including T-24 cells, treated with PAM were observed in vitro for changes in cell viability using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay. A viability reduction was detected in the various cancer cells after incubation in PAM. Furthermore, the study's results revealed that PAM was effective against cisplatin-resistant T-24 cells in vitro. In addition, a possible connection between HER expression and cell viability was sketched.

Biocompatibility and antibacterial activity of MgO/Ca3(PO4)2 composite ceramic scaffold based on vat photopolymerization technology

Recent advancements in medical technology and increased interdisciplinary research have facilitated the development of the field of medical engineering. Specifically, in bone repair, researchers and potential users have placed greater demands on orthopedic implants regarding their biocompatibility, degradation rates, antibacterial properties, and other aspects. In response, our team developed composite ceramic samples using degradable materials calcium phosphate and magnesium oxide through the vat photopolymerization (VP) technique. The calcium phosphate content in each sample was, respectively, 80 %, 60 %, 40 %, and 20 %. To explore the relationship between the biocompatibility, antibacterial activity, and MgO content of the samples, we cultured them with osteoblasts (MC3T3-E1), Escherichia coli (a gram-negative bacterium), and Staphylococcus aureus (a gram-positive bacterium). Our results demonstrate that as the MgO content of the sample increases, its biocompatibility improves but its antibacterial activity decreases. Regarding the composite material samples, the 20 % calcium phosphate content group exhibited the best biocompatibility. However, after 0.5 h of co-cultivation, the antibacterial rates of all groups except the 20 % calcium phosphate content group co-cultured with S. aureus exceed 80 %. Furthermore, after 3 h, the antibacterial rates against E. coli exceed 95 % in all groups. This is because higher levels of MgO correspond to lower pH values and Mg2+ concentrations in the cell and bacterial culture solutions, which ultimately promote cell and bacterial proliferation. This elevates the biocompatibility of the samples, albeit at the expense of their antimicrobial efficacy. Thus, modulating the MgO content in the composite ceramic samples provides a strategy to develop gradient composite scaffolds for better control of their biocompatibility and antibacterial performance during different stages of bone regeneration.

Aggregation of Thermoresponsive Polymethacrylates in a Dulbecco's Modified Eagle Medium and Its Salts.

The thermal behavior and aggregation process of the poly(N-isopropyl acrylamide), poly[oligo(ethylene glycol) methyl ether methacrylate], and poly[(2-hydroxyethyl methacrylate)-co-oligo(ethylene glycol) methyl ether methacrylate] thermoresponsive polymers were studied in a commonly used Dulbecco's Modified Eagle Medium (DMEM) cell culture medium and solutions of its individual components in the same concentration as found in DMEM. All studied copolymers exhibited an unexpected transmittance profile in the DMEM. During heating above the cloud point temperature (TCP), the polymers additionally aggregated, which led to the formation of their precipitates. The behavior of the polymers was further studied to evaluate how individual salts affected the transition temperature, size (Dh), and stability of the polymer particles. Organic additives, such as amino acids and glucose, had a significantly lesser impact on the thermoresponsive aggregation of the polymers than inorganic ones. Changes to the TCP were small and the formation of precipitates was not observed. The presence of small amounts of amino acids caused a decrease in the polymer aggregate sizes. Obtained results are of utmost importance in thermoresponsive drug nanocarrier studies.

Liquid Metal pH Morphology Sensor Used for Biological Microenvironment Detection.

pH is one of the important parameters of a biological microenvironment, which is closely related to cell growth, development, vitality, division, and differentiation. Monitoring the pH of a microenvironment is helpful to monitor the cell metabolism as well as to understand the cellular life cycle. The sensitivity of liquid metals (LMs) to hydrogen ions has aroused our interest. Here, we propose a novel but facile pH sensor using liquid gallium (LM for short) droplet morphological change as the readout. The pH sensing characteristics of the LM droplet were examined, especially the shape response. LM can form solid native oxide skin rapidly in oxygenated solution, and the oxide layer will be removed in acidic or alkaline solutions, which will cause a great change in surface tension. The phenomenon is the change of LM morphology from macroscopic observation. We explored the electrochemical characteristics of LM at different pH values, explained the mechanism of surface change, and calibrated the relationship curve between LM morphology and pH and the interference of impurity ions on the sensor. Finally, we proposed a detection algorithm for the LM pH morphology sensor and tried to automatically detect pH with a mobile app, which was applied to the pH detection of cell culture solution. We believe that the response characteristics of LM to hydrogen ions have great potential in microenvironment detection.

A Stream of Cell Culture and Process Liquid Solutions

Genetic Engineering & Biotechnology NewsVol. 42, No. 5 DepartmentsA Stream of Cell Culture and Process Liquid SolutionsGeminiBio, a reliable supplier of reagents, is adding new products, branching into instrumentation, and expanding its U.S.-based facilitiesGail DuttonGail DuttonSearch for more papers by this authorPublished Online:10 May 2022https://doi.org/10.1089/gen.42.05.05AboutSectionsView articleView Full TextPDF/EPUB Permissions & CitationsPermissionsDownload CitationsTrack CitationsAdd to favorites Back To Publication ShareShare onFacebookTwitterLinked InRedditEmail View articleFiguresReferencesRelatedDetails Volume 42Issue 5May 2022 Information© 2022 by GEN PublishingTo cite this article:Gail Dutton.A Stream of Cell Culture and Process Liquid Solutions.Genetic Engineering & Biotechnology News.May 2022.16-17.http://doi.org/10.1089/gen.42.05.05Published in Volume: 42 Issue 5: May 10, 2022PDF download

Technology roadmap for the development of a 3D cell culture workstation for a biomedical industry startup

This paper is a first attempt to establish how technology roadmapping, commonly reserved for established organizations, can be adapted for the characteristics and limitations of a startup company. This work was performed in collaboration with a newly founded company to create a technology roadmap for devising a 3D cell culture solution for the fields drug discovery and in vitro disease models. Currently, published literature focuses mainly on the methodology and the application in mature companies and industries and overlooking novel technological multinationals. This paper on the other hand, provides a case of implementing a technology roadmap for a start up technology building product based on an emerging technology in an emerging industry. The paper outlines how it was adopted and implemented successfully in a startup company. The roadmapping process was intended to create a strategic plan that can illustrate to the investors how the acquired resources will be used to develop the product. Additionally, it aimed to assist the company in disclosing diverse gaps and prioritizing efforts and resources to face uncertainty while developing the product and the company. The widespread use of advanced manufacturing has reached the drug discovery and research domain. There is an emerging market with no dominant design; currently, no solution addresses the requirements of the research and industry segments.

A Magnetocatalytic Propelled Cobalt–Platinum@Graphene Navigator for Enhanced Tumor Penetration and Theranostics

A Magnetocatalytic Propelled Cobalt–Platinum@Graphene Navigator for Enhanced Tumor Penetration and Theranostics

Exploring the degradation behavior of MgXAg alloys by in vitro electrochemical methods

Magnesium as biodegradable biomaterial could serve as bone augmentation material in implant dentistry. The knowledge about the predictability of the biodegradation process is essential as this process needs to go hand in hand with the formation of new bone to gradually replace the augmentation material. Therefore, this work aimed to assess if the electrochemistry (EC) measurements of the corrosion process correlate with the surface features at various time points during the surface degradation, in order to describe the degradation process of Mg and Mg alloys more reliably, under the assumption that differences in EC behavior can be detected and related to specific patterns on the surface.In this test setup, a special optical chamber was used for electrochemical measurements on Mg and Mg-alloys (Mg2Ag, Mg4Ag, and Mg6Ag). Specimens were investigated using different circulating cell culture solutions as electrolytes, these were minimum essential medium (MEM), Hank's Balanced Salt Solution (HBSS), and MEM+ (MEM with added sodium hydrogen carbonate) at 37 °C. Open circuit potential measurements (OCP) over 30 min followed by cyclic polarization were performed. The electrochemistry data, including OCP, exchange current density and corrosion potential, were compared with visible changes at the surface during these treatments over time. The results show that the addition of silver (Ag) leads to a “standardization” of the degradation regardless of the selected test medium. It is currently difficult to correlate the visible microscopic changes with the data taken from the measurements. Therefore, further investigations are necessary.

Marine cyanobacterium Spirulina maxima as an alternate to the animal cell culture medium supplement

Serum is a stable medium supplement for in vitro cell culture. Live cells are used in stem cell research, drug toxicity and safety testing, disease diagnosis and prevention, and development of antibiotics, drugs, and vaccines. However, use of serum in culture involves concerns such as an ethical debate regarding the collection process, lack of standardized ingredients, and high cost. Herein, therefore, we evaluated the possibility of using edible cyanobacterium (Spirulina maxima), which is a nutrient-rich, sustainable, and ethically acceptable source, as a novel substitute for fetal bovine serum (FBS). H460 cells were cultured to the 10th generation by adding a mixture of spirulina animal cell culture solution (SACCS) and FBS to the culture medium. Cell morphology and viability, cell cycle, apoptosis, proteomes, and transcriptomes were assessed. We observed that SACCS had better growth-promoting capabilities than FBS. Cell proliferation was promoted even when FBS was replaced by 50–70% SACCS; there was no significant difference in cell shape or viability. There were only slight differences in the cell cycle, apoptosis, proteomes, and transcriptomes of the cells grown in presence of SACCS. Therefore, SACCS has the potential to be an effective, low-cost, and eco-friendly alternative to FBS in in vitro culture.

Effectiveness of calcium hypochlorite, quaternary ammonium compounds, and sodium hypochlorite in eliminating vegetative cells and spores of Bacillus anthracis surrogate.

BackgroundThe spore-forming bacterium Bacillus anthracis causes anthrax, an often-fatal infection in animals. Therefore, a rapid and reliable strategy to decontaminate areas, humans, and livestock from B. anthracis is very critical.ObjectivesThe aim of this study was performed to evaluate the efficacy of sodium hypochlorite, calcium hypochlorite, and quaternary ammonium compound (QAC) sanitizers, which are commonly used in the food industry, to inhibit spores and vegetative cells of B. anthracis surrogate.MethodsWe evaluated the efficacy of sodium hypochlorite, calcium hypochlorite, and a QAC in inhibiting vegetative cells and spores of a B. anthracis surrogate. We treated a 0.1-mL vegetative cell culture or spore solution with 10 mL sanitizer. The samples were serially diluted and cultured.ResultsWe found that 50 ppm sodium hypochlorite (pH 7), 1 ppm calcium hypochlorite, and 1 ppm QAC completely eliminated the cells in vegetative state. Exposure to 3,000 ppm sodium hypochlorite (pH 7) and 300 ppm calcium hypochlorite significantly eliminated the bacterial spores; however, 50,000 ppm QAC could not eliminate all spores.ConclusionsCalcium hypochlorite and QAC showed better performance than sodium hypochlorite in completely eliminating vegetative cells of B. anthracis surrogate. QAC was ineffective against spores of the B. anthracis surrogate. Among the three commercial disinfectants tested, calcium hypochlorite most effectively eliminated both B. anthracis vegetative cells and spores.

MiRNA-1 promotes pyroptosis of cardiomyocytes and release of inflammatory factors by downregulating the expression level of PIK3R1 through the FoxO3a pathway.

Pyroptosis can be seen in both physiological and pathological processes, and whether it plays a positive or negative role has not been fully clarified, especially in the study of cardiology, which deserves more attention. A model of cardiomyocyte Hypoxia/Reoxygenation was established to detect MicroRNA (miRNA) expression. The C57BL/6 mice and H9c2 cells were respectively treated with Ischemic/Reperfusion (I/R) and Hypoxia/Reoxygenation (H/R). Then, the concentrations of interleukin-1β (IL-1β) and IL-18 were detected in serum and culture medium supernatant, as well as the expression levels of FOXO3, Caspase1, and PIK3R1. In I/R group, the concentrations of IL-1β and IL-18 in the supernatant of H9c2 cell culture solution were significantly higher compared with the control group. Consistent with the supernatant concentrations of IL-1β and IL-18, IL-1β, and IL-18 were also increased at a high level in I/R group at the transcriptional level. Furthermore, the in vitro experiments showed that FOXO3 and Caspase1 were significantly upregulated in myocardial cells with the treatment of I/R group compared to sham group. Concerning pyroptosis, the expression of PIK3R1 was dramatically decreased and the expression of miRNA-1 was significantly increased in H9c2 cells. MiRNA-1 promoted pyroptosis of cardiomyocytes and release of inflammatory factors by downregulating the expression level of PIK3R1.

Preparation of Cobalt–Chromium Alloy Planting Materials and Its Application in the Treatment of Gum Restoration

The preparation of cobalt–chromium alloy materials by different processes was studied and used to repair the full crown of experimental dogs. First, the casting method combined with 3D printing to obtain a cobalt–chromium alloy test piece, and the mechanical and microscopic properties of the test piece were analyzed during the preparation process; And then, selective laser melting (SLM) was used to perform single-channel, single-layer, and multi-layer scanning on the specimens obtained based on this technology during the preparation process. Two kinds of samples were selected from the cobalt–chromium test pieces obtained by the casting method, which were CS01 and CS02. And then selected one kind of sample from the cobalt–chromium test piece obtained by the SLM method, which was SLM01. Firstly, the roughness and contact angle of the planting material were compared, and the saliva-streptococcus mutant cell culture solution was configured to compare the amount of cell adhesion on the surface of different planting materials. Finally, the planting materials were used to repair the dog's full crown and establish animal models. The expressions of Bcl-2, Bax, Ki67 and p53 genes in gingival cells before and after canine crown wearing were compared in gingival tissues. The test results show that the mechanical properties and microstructure of the cobalt–chromium alloy material obtained based on the casting method meet the requirements. Based on the cobalt–chromium test piece obtained by the SLM method, the process parameters were optimized so that the corresponding density of the test piece reached 98.3%; Comparing roughness and surface contact angle, there was no statistical difference between different planting materials (P &gt; 0.05 ). At the same time, the number of Streptococcus mutant adhered to the surface of different specimens was not statistically significant (P &gt; 0.05 ); In the expression of gingival tissue, the implant material was obtained based on the SLM01 test piece, which promoted the decrease of Ki67 gene expression (P &lt; 0.05 ), and also promoted the proliferation of other gingival cell genes (P &lt; 0.05 ).

MiR-423-5p may regulate ovarian response to ovulation induction via CSF1

BackgroundWe have previously shown that hsa-miR-423-5p expression in ovarian granulosa cells is decreased in high ovarian response populations. The objective of the present study was to find the target gene and mechanism for miR-423-5p involved in ovarian response regulation.Methods(a) TargetScan was used to predict the target gene of hsa-miR-423-5p. (b) A model for hsa-miR-423-5p overexpression or inhibition was constructed by transfecting KGN cells with lentivirus. CSF1 mRNA and protein expression and luciferase activity were measured. (c) The cell cycles of control and lentivirus treated KGN cells were analyzed. Western blot was used to measure the expression of CDKN1A in KGN cells. (d) The concentration of E2 in KGN cell culture medium were measured.Results(a) TargetScan revealed that the 3′ un-translated region of CSF1 matched 11 bases at the 5′ end of miR-423-5p, making it a likely target gene. (b) Overexpression or inhibition of miR-423-5p were associated with respective decreases or increases in CSF1 expression (both mRNA and protein) (p < 0.05) and luciferase activity (p < 0.05). (c) When miR-423-5p expression increased, the number of G0/G1 phase cells and the expression of CDKN1A protein increased while estradiol concentrations in the cell culture solution decreased (p < 0.05). However, when miR-423-5p expression decreased, the number of S phase cells increased and E2 concentrations increased while the expression of CDKN1A protein decreased (p < 0.05).ConclusionsColony stimulating factor 1 is a target gene of miR-423-5p and that it may regulate ovarian response to ovulation induction by affecting granulosa cells proliferation and estrogen secretion.

Fabrication of Poly-Pyrrole Membrane Actuator for Cell Stimulation

Micro-actuators are used for mechanical stimulation of cultured cells in regenerative medicine and are critical components of biosensors. In this study, electrochemical polymerization is utilized to fabricate a film of poly-pyrrole (PPy) with a thickness of 10 μm. This film is peeled off from a working electrode substrate and subsequently laminated with a polydimethylsiloxane (PDMS) membrane containing holes of diameter of 5 mm. The assembled PPy film forms a membrane of PPy that can be used as a micro-actuator. This membrane is deflected upward via the application of voltages of −0.2, −0.4, −0.6, −0.8, and −1.0 V for 120 s in either NaDBS solution or cell culture solution. The primary response was an expansion in the in-plane direction with the absorption of ions in the electrolyte solution. The deflection increases with the duration of the applied voltage. Moreover, the maximum deflection that increases with the applied voltage reaches 540 μm at −1.0 V in the NaDBS solution. In the cell culture solution, the maximum deflection is approximately 400 μm for an applied voltage of −1.0 V. When the PPy membrane actuator was used in the culture solution, the time constant was 20 s to reach 63.2% of the maximum deflection. During operation, a voltage with a rectangular form and a period of 40 s was periodically applied. The operation of the PPy membrane actuator was repeated 90 times or more, although the deflection of the membrane had slight attenuation during the cycle of applied voltage. The PPy membrane exhibited adequate adhesiveness for cultured C2C12 cells. They adhered to the PPy surface and stretching of their pseudopods was observed. These cells are additionally cultured on the PPy membrane actuator. When a voltage is applied, the membrane actuator is operable while supporting cultured C2C12 cells. These cells are mechanically and electrically stimulated on the membrane that functions as a cell stimulation device.

Hepatoprotective effect of meal replacement seeds juice based on sweet potato\xa0(MRSJ) against CCl4-induced cytotoxicity in HepG2 cells

A sweet potato-based Meal Replacement Seeds Juice (MRSJ) was developed by mixing sweet potatoes and carrots with four types of seeds. Consuming the MRSJ rather than the whole vegetables or whole seeds improved digestive function, proving that it is suitable for the elderly. Its rich composition of minerals, vitamins, and unsaturated fatty acids implicates it as an excellent nutrient source. Notably, the ethyl acetate fraction of MRSJ contains abundant phenolics. The antioxidant activity assays showed that these phenolics have high radical scavenging activity, reducing power, and antioxidant capacity similar to l-ascorbic acid. The ethyl acetate fraction exerted protective effects against CCl4- or H2O2-induced hepatotoxicity via DNA protection, lipid accumulation inhibition, and cell protection, wherein ALT and AST activities in the cell culture solution decreased significantly. These findings suggest that MRSJ consumption may protect against liver diseases. Moreover, MRSJ as an excellent nutrient source may be developed as an age-neutral food.