Culture Material Research Articles

Comparative Study of Basement-membrane Matrices for Human Stem Cell Maintenance and Intestinal Organoid Generation.

Extracellular matrix (ECM) plays a critical role in cell behavior and development. Organoids generated from human induced pluripotent stem cells (hiPSCs) are in the spotlight of many research areas. However, the lack of physiological cues in classical cell culture materials hinders efficient iPSC differentiation. Incorporating commercially available ECM into stem cell culture provides physical and chemical cues beneficial for cell maintenance. Animal-derived commercially available basement membrane products are composed of ECM proteins and growth factors that support cell maintenance. Since the ECM holds tissue-specific properties that can modulate cell fate, xeno-free matrices are used to stream up translation to clinical studies. While commercially available matrices are widely used in hiPSC and organoid work, the equivalency of these matrices has not been evaluated yet. Here, a comparative study of hiPSC maintenance and human intestinal organoids (hIO) generation in four different matrices: Matrigel (Matrix 1-AB), Geltrex (Matrix 2-AB), Cultrex (Matrix 3-AB), and VitroGel (Matrix 4-XF) was conducted. Although the colonies lacked a perfectly round shape, there was minimal spontaneous differentiation, with over 85% of the cells expressing the stem cell marker SSEA-4. Matrix 4-XF led to the formation of 3D round clumps. Also, increasing the concentration of supplement and growth factors in the media used to make the Matrix 4-XF hydrogel solution improved hiPSC expression of SSEA-4 by 1.3-fold. Differentiation of Matrix 2-AB -maintained hiPSC led to fewer spheroid releases during the mid-/hindgut stage compared to the other animal-derived basement membranes. Compared to others, the xeno-free organoid matrix (Matrix 4-O3) leads to larger and more mature hIO, suggesting that the physical properties of xeno-free hydrogels can be harnessed to optimize organoid generation. Altogether, the results suggest that variations in the composition of different matrices affect stages of IO differentiation. This study raises awareness about the differences in commercially available matrices and provides a guide for matrix optimization during iPSC and IO work.

Metallographic study of non-ferrous metal products from the Maray 1 site of the Baitovo Cul-ture in the Lower Ishim River region

The article examines non-ferrous metal products from the Baitovo Culture materials of the Early Iron Age hillfort of Marai 1. This settlement is located in the forest-steppe of the Ishim River region of Western Siberia. The upper building hori-zon marks the settlement of the Baitovo Culture dating to the 4th–2nd centuries BC. Metallographic analysis of 28 items was carried out using the Axio Observer D1m microscope (Zeiss). As a result, eight technological schemes characteristic for the metalworking activities of the Baitovo population have been identified. The study shows that prevailing were smithing tech-nologies in different temperature regimes for the production of non-ferrous metal objects (16 specimens, 57.2 %). The pref-erence was given to hot forging in the red-heat regime at 600–800°C (7 specimens, 25 %), cold forging with annealing (5 specimens, 17.8 %), as well as incomplete hot forging at 300–500°C (3 specimens, 10.7 %). Casting without further refine-ment was recorded in the microstructures of arrowheads, ingots and splashes (9 specimens, 14.2 %). Casting, followed by a slight degree of deformation (20–30 %), was noted in three cases. This scheme correlates with the metalworking traditions of the Itkul and Krasnoozerskaya Cultures, and it was dominant on the territory of the Tobol-Ishim region during the Early Iron Age. Such a low percentage of items related to it could be explained by the absence in Marai 1 of large items cast in moulds needing further refinement (knives, celts, etc.). A microstructural study confirmed the use of pure oxidized copper by the craftsmen of the Baitovo Culture, with characteristic inclusions of Cu–Cu2O eutectic, associated with the Ural production centres. Arrowheads and an ingot made of pure oxidized copper were probably a direct import from the territory of the Itkul Culture. Low-alloy tin and arsenic bronzes, coming from the metal production centres of Kazakhstan and Altai, were also used to the same extent for production of some tools and jewellery. The correlation between the type of product, composition of raw materials and technology of its manufacture has been revealed, which is most clearly manifested in arrowheads and awls. The use of high temperatures and significant degrees of compression (80–90 %) has been recorded in a number of cases, resulting in the appearance of rejects. Cracks of hot brittleness, as well as of cold brittleness, occurred in the process of technological cycle, carried out with an excess of the tensile strength of the metal.

Efforts to Improve Motivation and Learning Outcomes with the Use of Powerpoint Media


 
 
 The purpose of this study was to determine the use of powerpoint media to increase motivation and learning outcomes of art and culture with materials for drawing decorative flora and fauna for class VIIIA students of SMP Negeri 02 Padangsidimpuan in the second semester of the academic year 2022/2023. This research was conducted in the second semester of the academic year 2022/2023. The research subjects were 36 students of class VIIIA of SMP Negeri 02 Padangsidimpuan. With the research target is the motivation and learning outcomes of art and culture materials for drawing decorative flora and fauna. Tool to collect data by observation to determine student learning motivation, learning outcomes of art and culture carried out a performance drawing test. This study uses 3 data analyzes, namely initial conditions, cycle 1 data, and cycle 2 data analyzed using comparative description techniques followed by reflection. The method used is to form large groups, small groups using powerpoint media are carried out by drawing performance tests to determine the increase in learning outcomes. This research model uses 4 stages of action: planning, action, observation, and reflection. Based on the results of the study, it was concluded that there had been an increase in the motivation indicator data for cycle 1, indicator 1) 81%, 2) 82%, 3) 87, cycle 2 increased to indicator 1) 83%, 2) 84%, 3) 90%. While the data on learning outcomes in cycle 1, the lowest score was 70, the highest score was 87, the average value was 77, there were 2 students who had not completed, cycle 2 the lowest score was 76, the highest score was 90, the average value was 79, and all the students completed.
 
 

Secondary Publication: Proposal for Points of Consideration for Pluripotent Stem Cell Culture

Human pluripotent stem cells, such as human embryonic stem cells and human induced pluripotent stem cells, are used in basic research and various applied fields, including drug discovery and regenerative medicine. Stem cell technologies have developed rapidly in recent years, and the supply of culture materials has improved. This has facilitated the culture of human pluripotent stem cells and has enabled an increasing number of researchers and bioengineers to access this technology. At the same time, it is a challenge to share the basic concepts and techniques of this technology among researchers and technicians to ensure the reproducibility of research results. Human pluripotent stem cells differ from conventional somatic cells in many aspects, and many points need to be considered in their handling, even for those experienced in cell culture. Therefore, we have prepared this proposal, “Points of Consideration for Pluripotent Stem Cell Culture,” to promote the effective use of human pluripotent stem cells. This proposal includes seven items to be considered and practices to be confirmed before using human pluripotent stem cells. These are laws/guidelines and consent/material transfer agreements, diversity of pluripotent stem cells, culture materials, thawing procedure, media exchange and cell passaging, freezing procedure, and culture management. We aim for the concept of these points of consideration to be shared by researchers and technicians involved in the cell culture of pluripotent stem cells. In this way, we hope the reliability of research using pluripotent stem cells can be improved, and cell culture technology will advance.

Tailoring the Degradation Time of Polycationic PEG-Based Hydrogels toward Dynamic Cell Culture Matrices.

Poly(ethylene glycol)-based (PEG) hydrogels provide an ideal platform to obtain well-defined and tailor-made cell culture matrices to enhance in vitro cell culture conditions, although cell adhesion is often challenging when the cells are cultivated on the substrate surface. We herein demonstrate two approaches for the synthesis of polycationic PEG-based hydrogels which were modified to enhance cell-matrix interactions, to improve two-dimensional (2D) cell culture, and catalyze hydrolytic degradation. While the utilization of N,N-(bisacryloxyethyl) amine (BAA) as cross-linker for in situ gelation provides degradable scaffolds for dynamic cell culture, the incorporation of short segments of poly(N-(3-(dimethylamino)propyl)acrylamide) (PDMAPAam) provides high local cationic charge density leading to PEG-based hydrogels with high selectivity for fibroblastic cell lines. The adsorption of transforming growth factor (TGF-β) into the hydrogels induced stimulation of fibrosis and thus the formation of collagen as a natural ECM compound. With this, these dynamic hydrogels enhance in vitro cell culture by providing a well-defined, artificial, and degradable matrix that stimulates cells to produce their own natural scaffold within a defined time frame.

Gender Violence in the Film Posesif by Gina S. Noer and its Relevance as Indonesian Language Teaching Material

Violence against women encompasses any kind of behavior that inflicts physical, sexual, or psychological damage against women. This study investigates the narrative structure of Gina S. Noer's film "Possessive," explores gender-based violence through the lens of feminist studies, and analyzes its significance to Indonesian language education. This research employs a descriptive-qualitative approach. We use an integrated and case study research methodology. This research investigates the film "Possessive" directed by Gina S. Noer, with a specific emphasis on the themes of gender-based violence and narrative structure. This research focuses on the portrayal of gendered violence against women in Gina S. Noer's film "Possessive" and its narrative structure. This inquiry used the film "Possessive" and literary literature. Information was gathered via library research. Data was confirmed using triangulation. Data analysis employs heuristic and hermeneutic approaches to interpret semiotic models. The research revealed that Gina S. Noer's film "Possessive" revolves on themes of love and violence, the narrative structure, the characters of Lala and Yudhis, as well as Lala's domestic and professional environment. Gina S. Noer's film "Possessive" depicts a total of five instances of verbal violence, three instances of psychological violence, three instances of physical violence, and three instances of power-based violence. (3) The findings of this research may be used to develop comprehensive Indonesian language teaching materials for high school students, including language, psychology, and culture.

Building a Better Defense: Expanding and Improving Natural Killer Cells for Adoptive Cell Therapy.

Natural killer (NK) cells have gained attention as a promising adoptive cell therapy platform for their potential to improve cancer treatments. NK cells offer distinct advantages over T-cells, including major histocompatibility complex class I (MHC-I)-independent tumor recognition and low risk of toxicity, even in an allogeneic setting. Despite this tremendous potential, challenges persist, such as limited in vivo persistence, reduced tumor infiltration, and low absolute NK cell numbers. This review outlines several strategies aiming to overcome these challenges. The developed strategies include optimizing NK cell expansion methods and improving NK cell antitumor responses by cytokine stimulation and genetic manipulations. Using K562 cells expressing membrane IL-15 or IL-21 with or without additional activating ligands like 4-1BBL allows "massive" NK cell expansion and makes multiple cell dosing and "off-the-shelf" efforts feasible. Further improvements in NK cell function can be reached by inducing memory-like NK cells, developing chimeric antigen receptor (CAR)-NK cells, or isolating NK-cell-based tumor-infiltrating lymphocytes (TILs). Memory-like NK cells demonstrate higher in vivo persistence and cytotoxicity, with early clinical trials demonstrating safety and promising efficacy. Recent trials using CAR-NK cells have also demonstrated a lack of any major toxicity, including cytokine release syndrome, and, yet, promising clinical activity. Recent data support that the presence of TIL-NK cells is associated with improved overall patient survival in different types of solid tumors such as head and neck, colorectal, breast, and gastric carcinomas, among the most significant. In conclusion, this review presents insights into the diverse strategies available for NK cell expansion, including the roles played by various cytokines, feeder cells, and culture material in influencing the activation phenotype, telomere length, and cytotoxic potential of expanded NK cells. Notably, genetically modified K562 cells have demonstrated significant efficacy in promoting NK cell expansion. Furthermore, culturing NK cells with IL-2 and IL-15 has been shown to improve expansion rates, while the presence of IL-12 and IL-21 has been linked to enhanced cytotoxic function. Overall, this review provides an overview of NK cell expansion methodologies, highlighting the current landscape of clinical trials and the key advancements to enhance NK-cell-based adoptive cell therapy.

Peptide‐Based Supramolecular Therapeutics for Fighting Major Diseases

AbstractPeptide‐based supramolecular therapeutics (PST) refer to those theranostic nanomedicines based on peptide or peptide derivatives self‐assembly that are constructed in vitro or in vivo. Benefiting from the inherent advantages of peptides, such as good biocompatibility, high bioactivity, and flexible tunability, the reported PST have shown excellent application prospects in combating many major diseases including cancer, cardiovascular disease, infectious disease, autoimmune disease, metabolic disease, and so on. Through thoughtful design, the PST can be fabricated in various forms, including self‐assembled precursor molecules, functional nano‐assemblies, and macroscopic large‐scale hydrogels. The involved peptides in the PST can play different roles, such as serving drug carriers for effective delivery, acting as scaffold materials for cell culture, and functioning as bioactive sequences for treating diseases. In this review, a comprehensive overview of traditional strategies is provided to build PST and emphasize their therapeutic applications for fighting major diseases that pose a threat to human life and health. Moreover, the main challenges of PST in systems design, therapeutic application, and clinical translation are also briefly discussed. It is hoped that this review can arouse much more attention to be paid for PST and promote its practical application in the biomedical field.

Contributions of Chinese hamster ovary cell derived extracellular vesicles and other cellular materials to hollow fiber filter fouling during perfusion manufacturing of monoclonal antibodies.

Hollow fiber filter fouling is a common issue plaguing perfusion production process for biologics therapeutics, but the nature of filter foulant has been elusive. Here we studied cell culture materials especially Chinese hamster ovary (CHO) cell-derived extracellular vesicles in perfusion process to determine their role in filter fouling. We found that the decrease of CHO-derived small extracellular vesicles (sEVs) with 50-200 nm in diameter in perfusion permeates always preceded the increase in transmembrane pressure (TMP) and subsequent decrease in product sieving, suggesting that sEVs might have been retained inside filters and contributed to filter fouling. Using scanning electron microscopy and helium ion microscopy, we found sEV-like structures in pores and on foulant patches of hollow fiber tangential flow filtration filter (HF-TFF) membranes. We also observed that the Day 28 TMP of perfusion culture correlated positively with the percentage of foulant patch areas. In addition, energy dispersive X-ray spectroscopy-based elemental mapping microscopy and spectroscopy analysis suggests that foulant patches had enriched cellular materials but not antifoam. Fluorescent staining results further indicate that these cellular materials could be DNA, proteins, and even adherent CHO cells. Lastly, in a small-scale HF-TFF model, addition of CHO-specific sEVs in CHO culture simulated filter fouling behaviors in a concentration-dependent manner. Based on these results, we proposed a mechanism of HF-TFF fouling, in which filter pore constriction by CHO sEVs is followed by cake formation of cellular materials on filter membrane.

Impact of Ionomycin on the Gene Expression of In vitro Fertilized Bovine Embryos

Background: The embryo genomic material used in this study came from an equal contribution of maternal and paternal genes. Gene expression during IVEP embryo development was controlled by several factors: the genetic makeup of the embryo, the epigenetic contributions and the culture materials. The aims of this research were to study the impact of ionomycin treatment on the sperm capacitation time and its impacts on the gene expression of some sensitive genes (Bcl-2, Bax, HSPB1, IGF1 and GDF9) during IVEP. Methods: The semen was placed in a sterile 15ml tube containing 5ml of capacitation medium and then split into three different experimental groups: the first group was the control; the second was the low-treated group, which contained sperm in a capacitation medium with 25 nM of ionomycin; the third group included 50 nM ionomycin. Result: The gene expression of BAX during early cleavage did not show any differences. However, in the late cleavage stage, the control group presented higher gene expression (P less than 0.05) than that in the two treated groups. The Bcl-2 genes of the control group showed higher gene expression (P less than 0.05) in both the early and late cleavage stages compared to the low-treated group. Gene expression of the HSP-1 gene was observed during early embryo development in the treated group (25 nM ionomycin). In conclusion, concentration 25 nM ionomycin showed higher gene expression in the early stages of embryo development also the best-quality embryos and gene expression results were found by studying gene expression. The transition from the maternal and paternal genome to the embryonic genome and gene expression during embryo development is controlled by several factors.

Design of dual peptide-conjugated hydrogels for proliferation and differentiation of human pluripotent stem cells

Completely synthetic cell cultivation materials for human pluripotent stem cells (hPSCs) are important for the future clinical use of hPSC-derived cells. Currently, cell culture materials conjugated with extracellular matrix (ECM)-derived peptides are being prepared using only one specific integrin-targeting peptide. We designed dual peptide-conjugated hydrogels, for which each peptide was selected from different ECM sites: the laminin β4 chain and fibronectin or vitronectin, which can target α6β1 and α2β1 or αVβ5. hPSCs cultured on dual peptide-conjugated hydrogels, especially on hydrogels conjugated with peptides obtained from the laminin β4 chain and vitronectin with a low peptide concentration of 200 μg/mL, showed high proliferation ability over the long term and differentiated into cells originating from 3 germ layers in vivo as well as a specific lineage of cardiac cells. The design of grafting peptides was also important, for which a joint segment and positive amino acids were added into the designed peptide. Because of the designed peptides on the hydrogels, only 200 μg/mL peptide solution was sufficient for grafting on the hydrogels, and the hydrogels supported hPSC cultures long-term; in contrast, in previous studies, greater than 1000 μg/mL peptide solution was needed for the grafting of peptides on cell culture materials.

Addition of Laponite to gelatin methacryloyl bioinks improves the rheological properties and printability to create mechanically tailorable cell culture matrices.

Extrusion-based bioprinting has gained widespread popularity in biofabrication due to its ability to assemble cells and biomaterials in precise patterns and form tissue-like constructs. To achieve this, bioinks must have rheological properties suitable for printing while maintaining cytocompatibility. However, many commonly used biomaterials do not meet the rheological requirements and therefore require modification for bioprinting applications. This study demonstrates the incorporation of Laponite-RD (LPN) into gelatin methacryloyl (GelMA) to produce highly customizable bioinks with desired rheological and mechanical properties for extrusion-based bioprinting. Bioink formulations were based on GelMA (5%-15% w/v) and LPN (0%-4% w/v), and a comprehensive rheological design was applied to evaluate key rheological properties necessary for extrusion-based bioprinting. The results showed that GelMA bioinks with LPN (1%-4% w/v) exhibited pronounced shear thinning and viscoelastic behavior, as well as improved thermal stability. Furthermore, a concentration window of 1%-2% (w/v) LPN to 5%-15% GelMA demonstrated enhanced rheological properties and printability required for extrusion-based bioprinting. Construct mechanical properties were highly tunable by varying polymer concentration and photocrosslinking parameters, with Young's moduli ranging from ∼0.2 to 75 kPa. Interestingly, at higher Laponite concentrations, GelMA cross-linking was inhibited, resulting in softer hydrogels. High viability of MCF-7 breast cancer cells was maintained in both free-swelling droplets and printed hydrogels, and metabolically active spheroids formed over 7 days of culture in all conditions. In summary, the addition of 1%-2% (w/v) LPN to gelatin-based bioinks significantly enhanced rheological properties and retained cell viability and proliferation, suggesting its suitability for extrusion-based bioprinting.

Measuring mechanical cues for modeling the stromal matrix in 3D cell cultures.

A breast-cancer tumor develops within a stroma, a tissue where a complex extracellular matrix surrounds cells, mediating the cancer progression through biomechanical and -chemical cues. Current materials partially mimic the stromal matrix in 3D cell cultures but methods for measuring the mechanical properties of the matrix at cell-relevant-length scales and stromal-stiffness levels are lacking. Here, to address this gap, we developed a characterization approach that employs probe-based microrheometry and Bayesian modeling to quantify length-scale-dependent mechanics and mechanical heterogeneity as in the stromal matrix. We examined the interpenetrating network (IPN) composed of alginate scaffolds (for adjusting mechanics) and type-1 collagen (a stromal-matrix constituent). We analyzed viscoelasticity: absolute-shear moduli (stiffness/elasticity) and phase angles (viscous and elastic characteristics). We determined the relationship between microrheometry and rheometry information. Microrheometry reveals lower stiffness at cell-relevant scales, compared to macroscale rheometry, with dependency on the length scale (10 to 100 μm). These data show increasing IPN stiffness with crosslinking until saturation (≃15 mM of Ca2+). Furthermore, we report that IPN stiffness can be adjusted by modulating collagen concentration and interconnectivity (by polymerization temperature). The IPNs are heterogeneous structurally (in SEM) and mechanically. Interestingly, increased alginate crosslinking changes IPN heterogeneity in stiffness but not in phase angle, until the saturation. In contrast, such changes are undetectable in alginate scaffolds. Our nonlinear viscoelasticity analysis at tumor-cell-exerted strains shows that only the softer IPNs stiffen with strain, like the stromal-collagen constituent. In summary, our approach can quantify the stromal-matrix-related viscoelasticity and is likely applicable to other materials in 3D culture.

Continuous flow delivery system for the perfusion of scaffold-based 3D cultures.

The paper-based culture platform developed by Whitesides readily incorporates tissue-like structures into laboratories with established workflows that rely on monolayer cultures. Cell-laden hydrogels are deposited in these porous scaffolds with micropipettes; these scaffolds support the thin gel slabs, allowing them to be evaluated individually or stacked into thick constructs. The paper-based culture platform has inspired many basic and translational studies, each exploring how readily accessible materials can generate complex structures that mimic aspects of tissues in vivo. Many of these examples have relied on static culture conditions, which result in diffusion-limited environments and cells experiencing pericellular hypoxia. Perfusion-based systems can alleviate pericellular hypoxia and other cell stresses by continually exposing the cells to fresh medium. These perfusion systems are common in microfluidic and organ-on-chip devices supporting cells as monolayer cultures or as 3D constructs. Here, we introduce a continuous flow delivery system, which uses parts readily produced with 3D printing to provide a self-contained culture platform in which cells in paper or other scaffolds are exposed to fresh (flowing) medium. We demonstrate the utility of this device with examples of cells maintained in single cell-laden scaffolds, stacks of cell-laden scaffolds, and scaffolds that contain monolayers of endothelial cells. These demonstrations highlight some possible experimental questions that can be enabled with readily accessible culture materials and a perfusion-based device that can be readily fabricated.

Identification of the Optimal Cultivation Period Required to Isolate Representatives of Mycobacterium abscessus Complex Isolated from Patients with Cystic Fibrosis.

In patients with cystic fibrosis (CF), representatives of the fast-growing Mycobacterium abscessus complex (MABSc) are often distinguished, but the culture of the material taken from such patients increases the growth time. We analyzed the terms of cultivation of MABSc representatives on dense nutrient media and also evaluated the productivity of a modified nutrient medium based on agar for the isolation of Burkholderia cepacia complex (BCC). Sixty-four strains of MABSc isolated from patients with CF and suspected tuberculosis were analyzed. The material from the patients was cultured on a universal chromogenic medium, 5% blood agar, yolk-salt agar, selective medium for isolation of BCC, and Löwenstein-Jensen medium. The cultures were incubated for 5 days (37°C, aerobic conditions), after for 23 days (28°C, aerobic conditions). The productivity of the developed nutrient medium was evaluated by the number of cells that gave visible growth after culturing 0.1 mL of a bacterial suspension of 103 CFU/mL. 76.8% of the strains grew in a 2-week period, and 23.2% of the strains were obtained at a later date from 18 to 28 days (average: 21.23 days). The modified medium with a concentration of 240 mg of iron (III) polymaltose hydroxide proved to be the most optimal for the isolation of MABSc. When using a chromogenic medium for culture material from patients with CF, it is necessary to extend incubation up to 28 days to increase the probability of MABSc isolation. The modified BCC medium showed a good selectivity result but required further investigation.

Pressing needs and recent advances to enhance production of embryos in vitro in cattle.

Embryo transfer in cattle is an increasingly important technique for cattle production. Full attainment of the benefits of the technology will depend on overcoming hurdles to optimal performance using embryos produced in vitro. Given its importance, embryo technology research should become a global research priority for animal reproduction science. Among the goals of that research should be developing methods to increase the proportion of oocytes becoming embryos through optimization of in vitro oocyte maturation and in vitro fertilization, producing an embryo competent to establish and maintain pregnancy after transfer, and increasing recipient fertility through selection, management and pharmacological manipulation. The embryo produced in vitro is susceptible to epigenetic reprogramming and methods should be found to minimize deleterious epigenetic change while altering the developmental program of the resultant calf to increase its health and productivity. There are widening opportunities to rethink the technological basis for much of the current practices for production and transfer of embryos because of explosive advances in fields of bioengineering such as microfluidics, three-dimensional printing of cell culture materials, organoid culture, live-cell imaging, and cryopreservation.

Photocrosslinkable microgels derived from human platelet lysates: injectable biomaterials for cardiac cell culture.

Cardiovascular diseases are a major global cause of morbidity and mortality, and they are often characterized by cardiomyocytes dead that ultimately leads to myocardial ischemia (MI). This condition replaces functional cardiac tissue with fibrotic scar tissue compromising heart function. Injectable systems for the in situ delivery of cells or molecules to assist during tissue repair have emerged as promising approaches for tissue engineering, particularly for myocardial repair. Methacryloyl platelet lysates (PLMA) have been employed for constructing full human-based 3D cell culture matrices and demonstrated potential for xeno-free applications. In this study, we propose using PLMA to produce microparticles (MPs) serving as anchors for cardiac and endothelial cells and ultimately as injectable systems for cardiac tissue repair. The herein reported PLMA MPs were produced by droplet microfluidics and showed great properties for cell attachment. More importantly, it is possible to show the capacity of PLMA MPs to serve as cell microcarriers even in the absence of animal-derived serum supplementation in the culture media.

Development of Augmented Reality-Based Flashcard Media "Oreng Madureh" as an Introduction to Madurese Culture

The development of augmented reality (AR) based flashcard media "Oreng Madureh" has the potential to become an effective and interesting educational and cultural promotion tool with a combination of AR and Madurese culture. This research aims to develop learning media using AR-based flashcard media on introductory Madurese culture material which includes an introduction to traditional clothing, typical food and Madurese traditional houses. This development research uses the ADDIE model which consists of five stages, namely analysis, design, development, implementation and evaluation. Data collection used observation, questionnaires, interviews and documentation techniques for 50 children aged 4-6 years in 4 trial stages, namely material expert trials, learning media experts, small group and large group field trials. The data was analyzed quantitatively and qualitatively with material expert questionnaire trial results of 91.07%, learning media expert trial results of 92.64% with very feasible criteria, no need for revision, small group trial results of 87.14% and the results of the large group trial were 84.89% in the high category. These results indicate that this media is suitable for use in the learning process. The development of this media has the implication of being able to attract children to understand and appreciate the cultural diversity around them through interactive learning activities, as well as developing children's fine motor and cognitive skills.

Craniological and dental non-metric data on the problem of the interaction between the repre-sentatives of the Sargatka and Bolsherechenskaya Cultures of the forest-steppe of the south Western Siberia

In the present paper, fully based on previously published craniological and dental non-metric data, the origins of common morpho-logical elements in the compositions of representatives of the Sargatka and Bolsherechenskaya Cultures are investigated. An attempt is made to assess the available data with the aid of the integrated analysis, implying combined examination of craniological and dental non-metric data for each population. The previous studies of the craniological and dental non-metric data showed that the series are significantly distinct in the differentiation by the vector ‘west-east’, or, in other words, Caucasoidness-Mongoloidness. Despite this, their certain affinity was recorded repeatedly, both in the analysis of craniometric data as well as in the study of odontoscopic data, but no explanation of this peculiarity has yet been proposed. The craniological sample of the Sargatka Culture comprised 158 crania and the dental non-metric one — 424 crania. The boundaries of the areal of the Sargatka Culture encompass the forest-steppe zone of the south of Western Siberia: the basins of the rivers Tobol, Ishim, and Irtysh, and the Baraba forest-steppe. The examined materials were divided in four groups corresponding to these territories. The craniological sample of the Bolsherechenskaya Culture comprised 118 crania, and the dental non-metric data amounted to 326 crania. The Bolsherechenskaya Culture materials originate from 11 burial grounds in the territory of the Novosibirsk Ob basin. All data were previously published. In the results, the population interaction of the bearers of the Sargat and Belsherechenskaya Cultures has been recorded, in which some representatives of the Kamenskaya Culture of the forest-steppe Altai were also involved, which is confirmed by archaeological data. However, it should be emphasized that this interaction was not so wide-scale. The Sargatka series, being located near to the Bolsherechenskaya ones, are also located in the same field with autochthonous Siberian groups. This becomes the evidence of the fact that not only the interaction in the Early Iron Age binds together the representa-tives of the Sargatka and Bolsherechenskaya Cultures. The fact is that the specificity of the anthropological composition of the Bolsherechenskaya people is directly related to the most ancient autochthonous Siberian populations, whereas the specific features of the Sargatka people — to southwestern migrants. However, an important result of the conducted analysis is that the composition of the Sargatka series retains the specific elements which are characteristic of the autochthonous populations of Western Siberia and which do not disappear under the influence of foreign migrant groups — this is the maturised structure of the lower molars.

Reflections on the melodramatic cultural matrix in anime

The article exposes a theoretical research on Cultural Studies and Reception Studies to, later, associate it with the research themes. Supported by theories, it’s observed the relationship between production and reception in a circular way, which incites questions about identities, hybridity, globalization, resignification, interaction, mediations and cultural matrices. With the exercise, it’s perceived that the use of melodramatic narrative strategies in productions such as the anime helps the dramatic construction and raises reflections on its transnational capacity, in such a way that melodrama can be understood as a transcultural matrix, as it crosses cultures, softens borders and contributes to the access and popularity of Japanese cultural products.