Culture Material Research Articles

Pumpkin seed proteins rival animal gelatin in increasing the cytoaffinity of edible microbeads for cell-based meat culture

Edible microbeads are hotly sought after for emerging cell-based meat culture but there are no major breakthroughs so far. Herein we report a functional edible microbead with alginate as core coated with pumpkin proteins as shell. Proteins from 11 plant-seeds were extracted and tested their cytoaffinity as gelatin replacer by grafting them on alginate microbeads and pumpkin seed protein coated microbeads shown the best performance in stimulating proliferation of C2C12 cells (by 17 folds in a week), 3T3-L1 adipocytes, chicken muscle satellite cells and primary porcine myoblast. The cytoaffinity of pumpkin seed protein coated microbeads comparable with that of animal gelatin microbeads. Protein sequencing analysis on pumpkin seed proteins found that it is rich in RGD tripeptide moiety, which are known to be enhance cytoaffinity. Our work advances our search for edible microbeads as ECM materials for cell-based meat culture.

The Building of Corporate Culture

This paper from the management science, economics, organization behavior science, philosophy, science of culture, psychology, sociology and systems theory and other aspects of the relevant theory of enterprise culture and its construction theory support. From the principle and characteristics of the corporate culture, the structure and function of enterprise culture, operation platform, enterprise culture evolution mechanism, operation path and so constructed theory analysis framework of enterprise culture. Based on the evaluation of elements from the spirit of enterprise culture, enterprise system culture, enterprise behavior culture and enterprise material culture, analyzes the effect of enterprise culture construction, on the basis of scientific, completeness, forward-looking, practical principles, established the effect of enterprise culture construction comprehensive evaluation model. The paper has important reference value for the healthy development of the enterprises of our country, also has certain reference significance to the academic research in the relevant field.

IMAGE OF WARTIME UKRAINE IN THE BRITISH AND AMERICAN MEDIA DISCOURSES

The article centers on the image of wartime Ukraine in media discourses of Great Britain and the USA. The publications, broadcasts and messages concerning Ukraine reveal not only the factual events or phenomena, pertaining to the people of Ukraine or their culture, but also cultural matrices and beliefs that determine how representatives of different countries understand and interpret the world. Media-driven discourses analysed in the article include various outlets that people often choose on the every-day basis: newspapers, magazines, radio, television, and the Internet.

Production of passion fruit virus-free in vitro shoots by apical meristem culture

Passion fruit is a high economic value crop. Currently, the cultivated area of passion fruit in Lam Dong is decreasing due to the spread of viral diseases. The main aim of this study is to generate disease-free in vitro shoots for propagation purposes. The apical shoots of purple (Passiflora edulis Sims.) and yellow (Passiflora edulis f. flavicarpa) passion fruit cultivars collected at Dam Rong (T1 and V1) and Duc Trong (T2 and V2) districts, without surface sterilisation, were used as materials for apical meristem culture. The apical meristems (0.2 mm) of passion fruit plants were cultured on MS medium supplemented with 1.0 mg/l BA. After 4 weeks of culture, the survival and regeneration rate reached over 45.0%. Single shoots with green leaves were formed and regenerated after 10 weeks of culture. These regenerated shoots were tested for the presence of 3 viral groups (CMV, ToRSV and Potyvirus) by RT-PCR. The results showed that only 1 of 4 tested samples (V1) was infected with Potyvirus. Thus, these virus-free in vitro shoots will be used as materials for propagation to meet the requirement for disease-free purple and yellow passion fruit plants.

Stiffness-Modulation of Collagen Gels by Genipin-Crosslinking for Cell Culture.

The stiffness of extracellular matrices (ECMs) is critical for cellular functions. Therefore, modulating the stiffness of ECMs in vitro is necessary to investigate the role of stiffness in cellular phenomena. Collagen gels are widely used for cell culture matrices in vitro. However, modulation of the stiffness in collagen gels for cell culture is challenging owing to the limited knowledge of the method to increase the stiffness while maintaining low cytotoxicity. Here, we established a novel method to modulate collagen gel stiffness from 0.0292 to 12.5 kPa with low cytotoxicity. We prepared collagens with genipin, a low-cytotoxic crosslinker of amines, at different concentrations and successfully modulated the stiffness of the gels. In addition, on 10 mM genipin-mixed collagen gels (approximately 12.5 kPa), H1299 human lung cancer cells showed spreading morphology and nuclear localization of yes-associated protein (YAP), typical phenomena of cells on stiff ECMs. Mouse mesenchymal stromal cells on 10 mM genipin-mixed collagen gels differentiated to vascular smooth muscle cells. On the other hand, the cells on 0 mM genipin-mixed collagen gels (approximately 0.0292 kPa) differentiated to visceral smooth muscle cells. Our new method provides a novel way to prepare stiffness-modulated collagen gels with low cytotoxicity in cell culture.

Toxicokinetics of a Single Oral Dose of OTA on Dezhou Male Donkeys.

Ochratoxin (OTA) is widely present in a wide range of foods and feeds, causing adverse effects on animals and humans. This study aims to explore the toxicokinetics of OTA-contaminated materials on the Dezhou male donkey. Donkeys received a single orally dose of 2500 μg OTA/kg BW, obtained from Aspergillus ochraceus culture material. The concentrations of OTA in plasma collected at 0, 5, 10, 15, 20, 30, 45 min, and at 1, 1.5, 2, 3, 6, 9, 12, 24, 48, 72, 96 and 120 h were detected by HPLC. OTA eliminated in urine and feces were quantified at 6-h intervals up to 24 h and then at 4-h intervals up to 120 h. The results suggested that the maximum concentration of OTA in plasma was observed at 12 h after administration, with a mean value of 10.34 μg/mL. The total excretion in both urine and feces was about 10% of the intake until 120 h.

Effect of Phthalates from Plastic Culture Materials on the Growth and Survival of African Catfish

Effect of Phthalates from Plastic Culture Materials on the Growth and Survival of African Catfish

Nature Returns: Dolphins and Dinosaurs. On Fake News, Photoshop Urban Legends and Memes during the COVID-19 Lockdown


 
 
 
 The article traces a narrative trail of social media posts and fake news in the early months of COVID-19 about how the reduction in pollution caused by the lockdown will allegedly have such a positive effect on the environment that nature will be restored. The study sheds light on this context and observes the change in rhetoric through the cultural matrices of various genres. It discusses whether an image is capable of telling a credible story. If it can, what is the relationship between context, truth and rhetoric and what role does humour play? In this article, however, the texts are accompanied by photographs depicting the events so it is easy to observe how visuality, which in fake news is part of the truth rhetoric, transforms into a vernacular critique of the truth itself. As to the material presented here, a contemporary legend researcher cannot ignore the legend motifs used in the viral visual narratives known as memes.
 
 
 

Stem cell culture and differentiation in 3-D scaffolds.

Conventional two-dimensional (2-D) cultivation are easy to utilize for human pluripotent stem (hPS) cell cultivation in standard techniques and are important for analysis or development of the signal pathways to keep pluripotent state of hPS cells cultivated on 2-D cell culture materials. However, the most efficient protocol to prepare hPS cells is the cell culture in a three dimensional (3-D) cultivation unit because huge numbers of hPS cells should be utilized in clinical treatment. Some 3-D cultivation strategies for hPS cells are considered: (a) microencapsulated cell cultivation in suspended hydrogels, (b) cell cultivation on microcarriers (MCs), (c) cell cultivation on self-aggregated spheroid [cell aggregates; embryoid bodies (EBs) and organoids], (d) cell cultivation on microfibers or nanofibers, and (e) cell cultivation in macroporous scaffolds. These cultivation ways are described in this chapter.

Advances in biomimetic hydrogels for organoid culture.

An organoid is a 3-dimensional (3D) cell culture system that mimics the structural and functional characteristics of organs, and it has promising applications in regenerative medicine, precision drug screening and personalised therapy. However, current culture techniques of organoids usually use mouse tumour-derived scaffolds (Matrigel) or other animal-derived decellularised extracellular matrices as culture systems with poorly defined components and undefined chemical and physical properties, which limit the growth of organoids and the reproducibility of culture conditions. In contrast, some synthetic culture materials have emerged in recent years with well-defined compositions, and flexible adjustment and optimisation of physical and chemical properties, which can effectively support organoid growth and development and prolong survival time of organoid in vitro. In this review, we will introduce the challenge of animal-derived decellularised extracellular matrices in organoid culture, and summarise the categories of biomimetic hydrogels currently used for organoid culture, and then discuss the future opportunities and perspectives in the development of advanced hydrogels in organoids. We hope that this review can promote academic communication in the field of organoid research and provide some assistance in advancing the development of organoid cultivation technology.

Polymerized Laminin-521: A Feasible Substrate for Expanding Induced Pluripotent Stem Cells at a Low Protein Concentration.

Laminins (LNs) play a central role in the self-assembly and maintenance of basement membranes and are involved in critical interactions between cells and other extracellular matrix (ECM) proteins. Among the defined, xeno-free ECM culture matrices, LNs-namely LN521-have emerged as promising coating systems for the large-scale expansion of induced pluripotent stem cells (iPSCs). The biologic activity of LNs is enhanced by their acidification-induced self-polymerization into a cell-associated network called polylaminin (polyLN), which can recapitulate the native-like polymeric array in a cell-free system. Here, we show for the first time to our knowledge that polyLN521 displays a native-like hexagonal-like structure and that, at basal and low concentrations, it permits the large-scale expansion of human iPSCs. Human iPSCs expanded with polyLN521 maintained the pluripotent state and showed no impairment of karyotype stability or telomere length. These results suggest that low-concentration polyLN521 is a stable and cost-effective coating for large-scale iPSC expansion.

Global communication of Chinese culture in the digital era: internal motivation and external challenges

The development of digital technology encourages culture to spread more inclusively and diversely around the world. On this basis, this article aims to reveal the changing trend and the dynamic mechanism behind Chinese cultural communication by studying the status quo of overseas communication in the media, education, and arts and the influence of Chinese culture in different regions. The research shows that the existence of cultural hegemony and unilateralism in Western countries and the incomplete of China's communication system have restricted the global communication of Chinese culture. Despite this, globalization and multi-polarization provide new external opportunities for Chinese cultural global communication. Also, the development of China's comprehensive strength and technology have laid the material foundation and broadened the channels for cultural communication. In the future, more revolutions in terms of cultural communication concepts, subjects, and matrices should be conducted to help Chinese culture be more widely recognized and accepted worldwide.

Development of “ARCIL” Media in Prehistoric Culture Materials for Junior High School Students

the purpose of this research and development is to produce learning media that are real, concrete, and interesting using augmented reality (AR) technology on learning material that contain pre-literate cultural products. This research and development uses ADDIE instructional design from the RnD method with five stages namely analysis, design, development, implementation, and evaluation. The data obtained is quantitative and qualitative data. Research instruments in the form of closed questionnaires with a level scale of 1-4 to obtain quantitative data and open questionnaires to obtain qualitative data about product usage advice. The results of the study show that, media eligibility based on the assessment of linguists obtained a 90.625%, material experts obtained a 94.12%, media experts obtained a 97.62%. Media practicality tests based on user responses by education practitioners gained a 90.91% and by students obtained a 90.58% score. Based on the results of the study, ARCIL learning media is very valid and practical to use in the learning process. There are several suggestions for further research, namely, (1) Adding other types of prehistorical culture results that are more diverse, (2) Provide variations in the form of other learning materials that are more diverse, (3) The addition of audio explanations so that students better understand the explanation of the material presented.

Model Pembelajaran Problem Based Learning sebagai Upaya Peningkatan Prestasi Belajar Mata Pelajaran Pendidikan Kewarganegaraan

Learning Pancasila and Citizenship Education in class XI IPA 3 Madrasah Aliyah Negeri 2 Surakarta has problems, including 1) student achievement results are still low 2) not using various learning methods and learning resources, so learning is less interesting and less fun; 3) students tend to be passive when learning takes place; 4) students' understanding of political culture material is still low. The research used classroom action research (CAR) consisting of 2 cycles. The purpose of this study was to improve students' understanding of the subject of political culture and learning achievement of Civics subjects in class XI IPA 3 MAN 2 Surakarta through the Problem Based Learning method. This research was conducted in class XI IPA 3 MAN 2 Surakarta in the academic year 2020/2021 by applying the Problem Based Learning method. Data collection for learning outcomes is done by observing learning activities, observing peers, and writing tests. Validation of this research data using the triangulation method by re-checking with colleagues in order to obtain accurate data. Data analysis in this study used comparative qualitative and quantitative descriptive data analysis. The performance indicator of PPKn MAN 2 Surakarta subjects is getting a score of > 75 and students complete 75% of the total number of students. The research was carried out in two cycles, each cycle consisting of four stages, namely the planning, implementation, observation, and reflection stages. The results of the study showed 1) The students completed Pre-cycle 17 students, increased to 25 students or 83.33% in the first cycle and increased to 31 students or 100% in the second cycle. Unfinished Students: from Pre-cycle 13 students, it decreased to 5 students in Cycle I and decreased to 0 in Cycle II. Highest Value: from Precycle 80, increased to 85 in cycle I and increased again to 90 in Cycle II. The lowest value of the Precycle was 70, remained at 70 in the first cycle, and increased again to 75 in the second cycle. The average value of the Pre-cycle class was 74.33, which increased to 76.33 in the first cycle and increased again to 78.17 in the second cycle. 2) After the application of the problem-based learning model, it can improve from the initial condition to the final condition after the first cycle and continue in the second cycle, there is a significant increase in the understanding of the material understanding of the nature of democracy.

Culture filtrate supplementation can be used to improve Mycobacterium tuberculosis culture positivity for spinal tuberculosis diagnosis.

Culture remains the gold standard to diagnose spinal tuberculosis (STB) despite the paucibacillary nature of the disease. Current methods can take up to 42 days to yield a result, delaying the ability to rapidly detect drug resistance. Studies have demonstrated the use of supplementation with culture filtrate (CF) from an axenic culture of Mycobacterium tuberculosis (Mtb) as a source of growth factors to improve culture rates. Our objective was to test a modified culture assay, utilizing CF supplemented media (CFSM), to improve culture positivity rates for suspected STB. Twelve patients with suspected STB were assessed by conventional culture (BACTEC™ MGIT 960), GeneXpert™ and standard histopathological examination. Spinal biopsies were taken from areas of diseased vertebral tissue or abscess, predetermined from MRI. Additional biopsies were obtained to assess CFSM for improved detection and faster culture of Mtb. All cases were diagnosed as STB and treated empirically for tuberculosis based on either bacteriological evidence (GeneXpert™, MGIT and/or CFSM positive), or based on clinical presentation. 5 specimens (45.45%) were positive for Mtb DNA as detected by GeneXpert™ and 1 specimen (8.33%) was cultured using MGIT (time to detection; 18 days). CFSM was able to culture 7 specimens (58.3%), with all CFSM positive specimens yielding a culture within 14 days. Two samples were positive only using the CFSM assay pointing to additional yield for diagnostic workup. Modification of standard culture can improve detection of Mtb and reduce time to positivity in individuals with STB where culture material is a requirement.

Soft surfaces induce neural differentiation via the neuron restrictive silencer factor

The physical and mechanical environment influence the differentiation and culture of stem cells. Early research in this field showed that mesenchymal stem cells (MSCs) adopt preferential lineages when cultured on surfaces of varying stiffness. Culture on very soft surfaces promoted neural differentiation. This effect has been observed in mesenchymal and embryonic stem cells and was shown to increase development of neural stem cells. However, providing further information on the molecular characterization of this phenomenon could help to guide the development of materials that enhance neural differentiation. Previously, our laboratory showed that chemical cues, forskolin and IBMX, induced neural differentiation of MSCs by downregulating the Neuron Restrictive Silencer Factor, leading to de-repression of neural gene expression. We sought to determine if the mechanism whereby soft surfaces induce neural differentiation could also involve NRSF function. We show that MSCs cultured on soft polydimethylsiloxane (PDMS) surfaces have reduced expression of NRSF as well as altered localization of NRSF. This suggests the modulation of transcription factors by surface substrate could be capitalized upon in designing materials for neural cell culture.

Engineered Tissue for Cardiac Regeneration: Current Status and Future Perspectives.

Heart failure (HF) is the leading cause of death worldwide. The most effective HF treatment is heart transplantation, the use of which is restricted by the limited supply of donor hearts. The human pluripotent stem cell (hPSC), including human embryonic stem cell (hESC) and the induced pluripotent stem cells (hiPSC), could be produced in an infinite manner and differentiated into cardiomyocytes (CMs) with high efficiency. The hPSC-CMs have, thus, offered a promising alternative for heart transplant. In this review, we introduce the tissue-engineering technologies for hPSC-CM, including the materials for cell culture and tissue formation, and the delivery means into the heart. The most recent progress in clinical application of hPSC-CMs is also introduced. In addition, the bottleneck limitations and future perspectives for clinical translation are further discussed.

Single-Walled Carbon Nanotubes as Fluorescent Probes for Monitoring the Self-Assembly and Morphology of Peptide/Polymer Hybrid Hydrogels.

Hydrogels formed via supramolecular self-assembly of fluorenylmethyloxycarbonyl (Fmoc)-conjugated amino acids provide excellent scaffolds for 3D cell culture, tissue engineering, and tissue recovery matrices. Such hydrogels are usually characterized by rheology or electron microscopy, which are invasive and cannot provide real-time information. Here, we incorporate near-infrared fluorescent single-walled carbon nanotubes (SWCNTs) into Fmoc-diphenylalanine hydrogels as fluorescent probes, reporting in real-time on the morphology and time-dependent structural changes of the self-assembled hydrogels in the transparency window of biological tissue. We further demonstrate that the gelation process and structural changes upon the addition of cross-linking ions are transduced into spectral modulations of the SWCNT-fluorescence. Moreover, morphological differences of the hydrogels induced by polymer additives are manifested in unique features in fluorescence images of the incorporated SWCNTs. SWCNTs can thus serve as optical probes for noninvasive, long-term monitoring of the self-assembly gelation process and the fate of the resulting peptide hydrogel during long-term usage.

Stimulus-Responsive, Gelatin-Containing Supramolecular Nanofibers as Switchable 3D Microenvironments for Cells.

Polymer- and/or protein-based nanofibers that promote stable cell adhesion have drawn increasing attention as well-defined models of the extracellular matrix. In this study, we fabricated two classes of stimulus-responsive fibers containing gelatin and supramolecular crosslinks to emulate the dynamic cellular microenvironment in vivo. Gelatin enabled cells to adhere without additional surface functionalization, while supramolecular crosslinks allowed for the reversible switching of the Young’s modulus through changes in the concentration of guest molecules in culture media. The first class of nanofibers was prepared by coupling the host–guest inclusion complex to gelatin before electrospinning (pre-conjugation), while the second class of nanofibers was fabricated by coupling gelatin to polyacrylamide functionalized with host or guest moieties, followed by conjugation in the electrospinning solution (post-conjugation). In situ AFM nano-indentation demonstrated the reversible switching of the Young’s modulus between 2–3 kPa and 0.2–0.3 kPa under physiological conditions by adding/removing soluble guest molecules. As the concentration of additives does not affect cell viability, the supramolecular fibers established in this study are a promising candidate for various biomedical applications, such as standardized three-dimensional culture matrices for somatic cells and the regulation of stem cell differentiation.

An Aquaponic System with Hydroponic Culture of Sweet Potato and Tilapia Culture

An integrated system with a hydroponic culture of sweet potato and tilapia culture was assessed as a new combination in aquaponics to investigate its potential and productivity. The first experiment was conducted inside a glasshouse using plastic tanks as water reservoirs to compare the growth and development of two sweet potato (Ipomoea batatas (L.) Lam.) varieties Beniazuma and purple sweet lord using hydroballs, granular plastics, cut rockwool, and rockwool blocks as supporting materials in hydroponic culture. The supporting material was placed inside plastic pots. Greater growth and development of the storage roots of sweet potatoes in both varieties were obtained in hydroballs. In the second experiment, four sweet potato varieties, Kokei 14, Suioh, Beniazuma, and Elegant summer, with hydroballs as the supporting material, were examined hydroponically with an integrated culture of tilapia fish in the pond. Two ponds were 7.7 m and 5.3 m, in length and width, respectively, and water depth was maintained at 0.38 cm. Styrofoam boxes with holes on the bottom side were used to culture sweet potato on the water surface. Nylon net containers filled with hydroballs were placed in the Styrofoam boxes. The variety Kokei 14 produced the greatest fresh and dry weights of storage roots and percent harvest index in both ponds, with and without fish. The number and diameter of the storage roots of Kokei 14 were higher. Tilapia fish were cultured in one of the two ponds and fed artificial feed. The average length and weight of tilapia fishes increased to 10 cm and 188 g, respectively, 143 days after culturing, and those were 1.8 and 6.9 times, respectively, greater than at the start. The integrated culture of sweet potato hydroponically on the water surface and tilapia in pond water increases the food production in a limited space and profitable.