Culture Platform Research Articles

Scalable production of uniform and mature organoids in a 3D geometrically-engineered permeable membrane

The application of organoids has been limited by the lack of methods for producing uniformly mature organoids at scale. This study introduces an organoid culture platform, called UniMat, which addresses the challenges of uniformity and maturity simultaneously. UniMat is designed to not only ensure consistent organoid growth but also facilitate an unrestricted supply of soluble factors by a 3D geometrically-engineered, permeable membrane-based platform. Using UniMat, we demonstrate the scalable generation of kidney organoids with enhanced uniformity in both structure and function compared to conventional methods. Notably, kidney organoids within UniMat show improved maturation, showing increased expression of nephron transcripts, more in vivo-like cell-type balance, enhanced vascularization, and better long-term stability. Moreover, UniMat’s design offers a more standardized organoid model for disease modeling and drug testing, as demonstrated by polycystic-kidney disease and acute kidney injury modeling. In essence, UniMat presents a valuable platform for organoid technology, with potential applications in organ development, disease modeling, and drug screening.

Bridging the Skills Gap Through Education Supply Chain to Meet Industry Requirements

In Bangladesh, a persistent mismatch exists between the skills fresh graduates possess and the competencies demanded by employers. This gap affects graduate employability and hampers economic growth. This article explores how key components— Program Curriculum Development (PCD), Instructor Portfolio Management (IPM), University Culture and Networking Platform (UCN), and University Facilities (UF)—can collectively address this issue. By integrating these elements into the educational supply chain, universities can produce skilled graduates who are better equipped to meet industry needs. The proposed framework offers a comprehensive approach to align academic outputs with labor market demands. The study highlights the importance of adapting curricula, engaging industry professionals in teaching, fostering strong university-industry connections, and investing in modern facilities to close the skills gap. This holistic approach ensures that graduates are employable, contributing to national development and societal betterment. The article concludes by emphasizing the need for further research on technological advancements and policy reforms to enhance higher education in Bangladesh.

Understanding Charitable Consumer Behavior on Digital Crowdfunding Platforms: Exploring the Influence of Cultural Values, Motivations, and Platform Attitudes

ABSTRACT Charitable crowdfunding on digital platforms has become a viable way for NGOs to raise funds. This study aims to investigate determinants of charitable behavior (CB) on digital crowdfunding platforms (DCPs), including national culture dimensions, motivations, crowdfunding platform attitude, and donors’ personal characteristics. Data were collected through a CAWI survey of 680 Amazon MTurk users from Europe, the Americas, and Southern Asia. Findings indicate that of the three national culture dimensions considered, CB on DCPs is positively associated with collectivism and negatively with uncertainty avoidance. The link between power distance and CB could be either positive, neutral or negative, due to the interactions with motivations for charity, uncertainty avoidance, and age. Extrinsic and intrinsic motivations do not show significant direct effects on CB but appear to exert their influence through interactions with other motivations, national culture dimensions and respondents’ attributes. People with higher incomes tend to donate more the better is their attitude toward the DCP. CB is stronger among donors more involved in voluntary activities and those living in the Americas as compared to Europe and Asia. The study addresses the knowledge gap concerning the antecedents of CB on DCPs and suggests how NGOs could improve their fundraising effectiveness.

Under the iceberg of “one-stop”: explore multi-actor engagement in one-stop digital cultural platforms through actor-network and commons theories

PurposeThis study investigates the roles and interactions of various actors of one-stop digital culture platforms (ODCPs) by applying actor-network theory (ANT).Design/methodology/approachData were collected from focus group interviews with 19 participants and over 20 policy texts related to the digitization of cultural services in China. Utilizing ANT as the analytical framework, we employed the grounded theory to analyze the dataset.FindingsThe empirical results indicate that: (1) governments, cultural institutions, enterprises and users each play dual roles, contributing to the functionality and development of ODCPs; (2) introduces the theory of commons to reconceptualize ODCPs as commons, highlighting the need for collective management and shared responsibility in the governance of ODCPs.Originality/valueThis study identifies and elaborates on the different roles of stakeholders and addresses conceptual ambiguities within ODCP projects. By introducing the theory of commons into the governance of ODCPs, this research provides new theoretical guidance for decision-makers and practitioners involved in ODCP projects.

An Innovative Perspective for Constructing Cultural and Creative e-Commerce Platforms: Integrating Insights from ICDT Theory and Brand Personality

Thriving in competitive business environments requires innovation, especially within cultural and creative-based e-commerce platforms. This study examines the application of two prominent theories, ICDT theory and brand personality, in assessing new business models. Focusing on 479 respondents from Taiwan, this research explores how the implementation of ICDT and brand personality in cultural and creative e-commerce influences customer behavior. Statistical analyses and the Smart PLS method are adopted to identify the significance of and relationships between the variables in the structural equation model. The profound influence of the ICDT model is evident in the emergence of virtual information space, virtual communication space, and virtual transaction space. Brand personality traits underscore the significance of intellectual excellence, M-type traits, seeking innovation and change, and stable reliability in shaping customer values. This research enriches the literature by emphasizing the essential role of innovative technologies and brand issues integrated with core product attributes for successful consumer acceptance in the fiercely competitive landscape of e-commerce platforms.

Mechanical memory based biofabrication of hierarchical elastic cardiac tissue.

Mimicking the multilayered, anisotropic, elastic structure of cardiac tissues for controlled guidiance of 3D cellular orientation is essential in designing bionic scaffolds for cardiac tissue biofabrication. Here, a hierarchically organized, anisotropic, wavy and conductive polycaprolactone (PCL)/Au scaffold was created in a facile fashion based on mechanical memory during fabrication. The bionic 3D scaffold shows good biocompatibility, excellent biomimetic mechanical properties that guide myoblast alignment, support the hyperelastic behaviour observed in native cardiac muscle tissue, and promote myotube maturation, which holds potential for cardiac muscle engineering and the establishment of an in vitro culture platform for drug screening.

Data-driven optimization of the in silico design of ionic liquids as interfacial cell culture fluids

ABSTRACT As an alternative to conventional plastic dishes, the interface between water-immiscible hydrophobic fluids, such as perfluorocarbons and silicones, permits cell adhesion and growth. Thus, it is expected to replace the petroleum-derived products in a sustainable society. However, most hydrophobic fluids are cytotoxic, which limits the range of mechanical and chemical cues exposed to the cells. Using a data-driven approach, this study aimed to identify non-cytotoxic ionic liquids (ILs) as fluid culture platforms to take advantage of their ‘designer’ nature for broadening the possible physicochemical ranges exposed to cells and their repeated use owing to their high heat stability before their biological applications. The new candidates within the readily synthesize ammonium-type ILs were identified through the active cycle of regression and a limited number of cytotoxicity tests. Structure – cytotoxicity analysis indicated that the presence of multiple long alkyl branches was critical for low cytotoxicity. Particularly, we successfully cultured human mesenchymal stem cells (hMSCs) at the trihexylethylammonium trifluoromethylsulfonylimide interface and repeated their use after solvent extraction and heat sterilization. This study identified non-cytotoxic ILs that fulfill plastics’ 3 R ( R educe, R ecycle, and R eplace) requirements and opens new avenues for hMSC fate manipulation through mechanotransduction.

Dynamic reciprocal interactions between activated T cells and tumor associated macrophages drive macrophage reprogramming and proinflammatory T cell migration within prostate tumor models

Tumor-associated macrophages (TAMs) have been implicated as a tumor microenvironment (TME) cell population, which may be playing a vital role in the inhibition of effective T cell responses in the prostate TME. In this manuscript, we leverage a novel microscale cell culture platform, known as Stacks, to investigate mono-, co-, and tri-culture TME models comprised of prostate tumor cell lines, primary macrophages, and autologous T cells from patients with prostate cancer. Through multiplexed analysis of these multi-cellular prostate tumor models, we capture a dynamic interaction between primary TAMs and activated T cells that resulted in reciprocal proinflammatory activation of both cell populations upon interaction. These findings suggest that activated T cells are capable of reprogramming immunosuppressive TAMs in the context of prostate tumor models and that TAM reprogramming may play a key supportive role in restoring proinflammatory T cell tumor responses in the prostate TME.

How to Realize the Collaborative Supply of Cultural Resource Big Data with Government Participation: Experiences from China

To foster the sustainable development of culture, particularly focusing on the preservation of cultural heritage, encompassing relics, intangible cultural heritage, and historical sites, China has launched a strategy for the digitalization of culture, with the goal of establishing a holistic national big data framework for cultural resources. To improve the efficiency of collaborative supply of cultural resource big data among various parties and to further advance the sustainable development of culture, this research has created a cooperative model that includes cultural institutions, a cultural resource big data service platform, and government participation. Our research findings, based on prospect theory and evolutionary game theory combined with Chinese practice, are presented below. (1) Various factors, including the coefficient of digital infrastructure empowerment, access charges for digital infrastructure, government penalties, and the probability of data leakage, have varying effects on the system in different states. (2) Once the industry has developed, the government can increase the impact of digital infrastructure empowerment to create stronger incentives, rather than relying solely on rewards or penalties. (3) When the value level of cultural resource big data is high, the benefit distribution coefficient does not affect the system evolution results. Finally, we offer practical insights for the government, cultural organizations, and cultural resource big data service platforms based on our research results. Our research offers Chinese insights for global cultural sustainable development.

Temporal evolution of mechanical properties in PDMS: A comparative study of elastic modulus and relaxation time for storage in air and aqueous environment

Polydimethylsiloxane (PDMS) is a soft, biocompatible polymer extensively employed in biomedical research, notable for its tunable mechanical properties achieved through cross-linking. While many studies have assessed the mechanical properties of PDMS utilizing macroscopic and microscopic methods, these analyses are often limited to freshly prepared samples. However, the mechanical properties of PDMS can be expected to change during prolonged exposure to water or air, such as interface polymer chain loosening or surface hardening, which are critical considerations in applications like cell culture platforms or microfluidic devices. This paper presents a comprehensive 10-day investigation of the evolution of PDMS surface mechanical properties through AFM-based nano-indentation. We focused on the most commonly utilized crosslinker-to-base ratios of PDMS, 1:10 (r10) and 1:20 (r20), under conditions of air and deionized water storage. For r10 samples, a hardening process was detected, peaking at 2.12 ± 0.35 MPa within five days for those stored in air and 1.71 ± 0.16 MPa by the third day for those immersed in water. During indentation, the samples displayed multiple contact points, suggesting the formation of distinct regions with varying mechanical properties. In contrast, r20 samples exhibited better stability, with an observed elastic modulus averaging 0.62 ± 0.06 MPa for air-stored and 0.74 ± 0.06 MPa for water-stored samples. Relaxation experiments, interpreted via the General Maxwell Model featuring two distinct component responses, a relatively consistent fast response τ1 (on the order of 10−1 s), and a more variable, slower response τ2 (on the order of 10 s), throughout the study period. The identification of two distinct relaxation times suggests the involvement of two disparate material property regimes in the relaxation process, implying changes in the surface material composition at the interface with air/water. These variations in mechanical properties could significantly influence the long-term functionality of PDMS in various biomedical applications.

Combining human liver ECM with topographically featured electrospun scaffolds for engineering hepatic microenvironment

Liver disease cases are rapidly expanding worldwide, and transplantation remains the only effective cure for end-stage disease. There is an increasing demand for developing potential drug treatments, and regenerative therapies using in-vitro culture platforms. Human decellularized extracellular matrix (dECM) is an appealing alternative to conventional animal tissues as it contains human-specific proteins and can serve as scaffolding materials. Herein we exploit this with human donor tissue from discarded liver which was not suitable for transplant using a synergistic approach to combining biological and topographical cues in electrospun materials as an in-vitro culture platform. To realise this, we developed a methodology for incorporating human liver dECM into electrospun polycaprolactone (PCL) fibres with surface nanotopographies (230–580 nm). The hybrid scaffolds were fabricated using varying concentrations of dECM; their morphology, mechanical properties, hydrophilicity and stability were analysed. The scaffolds were validated using HepG2 and primary mouse hepatocytes, with subsequent results indicating that the modified scaffolds-maintained cell growth and influenced cell attachment, proliferation and hepatic-related gene expression. This work demonstrates a novel approach to harvesting the potential from decellularized human tissues in the form of innovative in-vitro culture platforms for liver.

Hydrogel Applications in the Diagnosis and Treatment of Glioblastoma.

Glioblastoma multiforme (GBM), a common malignant neurological tumor, has boundaries indistinguishable from those of normal tissue, making complete surgical removal ineffective. The blood-brain barrier (BBB) further impedes the efficacy of radiotherapy and chemotherapy, leading to suboptimal treatment outcomes and a heightened probability of recurrence. Hydrogels offer multiple advantages for GBM diagnosis and treatment, including overcoming the BBB for improved drug delivery, controlled drug release for long-term efficacy, and enhanced relaxation properties of magnetic resonance imaging (MRI) contrast agents. Hydrogels, with their excellent biocompatibility and customizability, can mimic the in vivo microenvironment, support tumor cell culture, enable drug screening, and facilitate the study of tumor invasion and metastasis. This paper reviews the classification of hydrogels and recent research for the diagnosis and treatment of GBM, including their applications as cell culture platforms and drugs including imaging contrast agents carriers. The mechanisms of drug release from hydrogels and methods to monitor the activity of hydrogel-loaded drugs are also discussed. This review is intended to facilitate a more comprehensive understanding of the current state of GBM research. It offers insights into the design of integrated hydrogel-based GBM diagnosis and treatment with the objective of achieving the desired therapeutic effect and improving the prognosis of GBM.

Convenient rapid prototyping microphysiological niche for mimicking liver native basement membrane: Liver sinusoid on a chip

Liver is responsible for the metabolization processes of up to 90 % of compounds and toxins in the body. Therefore liver-on-a-chip systems, as an in vitro promising cell culture platform, have great importance for fundamental science and drug development. In most of the liver-on-a-chip studies, seeding cells on both sides of a porous membrane, which represents the basement membrane, fail to resemble the native characteristics of biochemical, biophysical, and mechanical properties. In this study, polycarbonate (PC) and polyethylene terephthalate (PET) membranes were coated with gelatin to address this issue by accurately mimicking the native basement membrane present in the space of Disse. Various coating methods were used, including doctor blade, gel micro-injection, electrospinning, and spin coating. Spin coating was demonstrated to be the most effective technique owing to the ability to produce thin gel thickness with desirable surface roughness for cell interactions on both sides of the membrane. HepG2 and EA.HY926 cells were seeded on the upper and bottom sides of the gelatin-coated PET membrane and cultured on-chip for 7 days. Cell viability increased from 90 % to 95 %, while apoptotic index decreased. Albumin secretion notably rose between days 1–7 and 4–7, while GST-α secretion decreased from day 1 to day 7. In conclusion, the optimized spin coating process reported here can effectively modify the membranes to better mimic the native basement membrane niche characteristics.

Barrier-free, open-top microfluidic chip for generating two distinct, interconnected 3D microvascular networks

Developing microphysiological cell culture platforms with a three-dimensional (3D) microenvironment has been a significant advancement from traditional monolayer cultures. Still, most of the current microphysiological platforms are limited in closed designs, i.e. are not accessible after 3D cell culture loading. Here, we report an open-top microfluidic chip which enables the generation of two sequentially loaded 3D cell cultures without physical barriers restricting the nurture, gas exchange and cellular communication. As a proof-of-concept, we demonstrated the formation of two 3D vasculatures, one in the upper and the other in the lower compartment, under three distinct flow conditions: asymmetric side-to-center, symmetric side-to-center and symmetric center-to-side. We used computational modelling to characterize initial flow pressures in cell culture compartments. We showed prominent vessel formation and branched vasculatures in upper and lower cell culture compartments with interconnecting, lumenized vessels with in vivo-relevant diameter in all flow conditions. With advanced image processing, we quantified and compared the overall vascular network volume and the total length formed in asymmetric side-to-center, symmetric side-to-center and symmetric center-to-side flow conditions. Our results indicate that the developed chip can house two distinct 3D cell cultures with merging vessels between compartments and by providing asymmetric side-to-center or symmetric center-to-side flow vascular morphogenesis is enhanced in terms of overall network length. The developed open-top microfluidic chip may find various applications in generation of tissue-specific 3D-3D co-cultures for studying cellular interactions in vascularized tissues and organs.

Hydrogel microsphere stem cell encapsulation enhances cardiomyocyte differentiation and functionality in scalable suspension system

A reliable suspension-based platform for scaling engineered cardiac tissue (ECT) production from human induced pluripotent stem cells (hiPSCs) is crucial for regenerative therapies. Here, we compared the production and functionality of ECTs formed using our scaffold-based, engineered tissue microsphere differentiation approach with those formed using the prevalent scaffold-free aggregate platform. We utilized a microfluidic system for the rapid (1 million cells/min), high density (30, 40, 60 million cells/ml) encapsulation of hiPSCs within PEG-fibrinogen hydrogel microspheres. HiPSC-laden microspheres and aggregates underwent suspension-based cardiac differentiation in chemically defined media. In comparison to aggregates, microspheres maintained consistent size and shape initially, over time, and within and between batches. Initial size and shape coefficients of variation for microspheres were eight and three times lower, respectively, compared to aggregates. On day 10, microsphere cardiomyocyte (CM) content was 27 % higher and the number of CMs per initial hiPSC was 250 % higher than in aggregates. Contraction and relaxation velocities of microspheres were four and nine times higher than those of aggregates, respectively. Microsphere contractile functionality also improved with culture time, whereas aggregate functionality remained unchanged. Additionally, microspheres displayed improved β-adrenergic signaling responsiveness and uniform calcium transient propagation. Transcriptomic analysis revealed that while both microspheres and aggregates demonstrated similar gene regulation patterns associated with cardiomyocyte differentiation, heart development, cardiac muscle contraction, and sarcomere organization, the microspheres exhibited more pronounced transcriptional changes over time. Taken together, these results highlight the capability of the microsphere platform for scaling up biomanufacturing of ECTs in a suspension-based culture platform.

Recording and Interpretation of Active Calcium Transients in Induced Pluripotent Stem Cell-Derived Cardiomyocytes.

Calcium plays a pivotal role in the excitation-contraction coupling process in cardiomyocytes, a critical multi-parametric event leading to rhythmic contraction. Over the past few decades, calcium signaling in cardiomyocytes has been extensively studied in cardiovascular sciences. However, a standard methodology is needed not only to trace the calcium within cells but also to remove signal processing biases and to accurately interpret the features of calcium transient signals in relation to cardiomyocyte electrophysiology. This article outlines the use of genetically encoded calcium indicator (GCaMP) human-induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) to record calcium transients. These cells express a green fluorescent signal when calcium binds to intracellular calmodulin, a key regulator of calcium signaling. The extraction and processing of calcium transient waveforms are performed using ImageJ and MATLAB software. Key features of these waveforms are then identified and categorized based on their physiological relevance to cardiomyocyte function. Additionally, this work includes a Support Protocol for the successful replating of cardiomyocytes onto non-traditional culture platforms, such as metallic sensors and polymer-based substrates, to facilitate data multiplexing. The three Basic Protocols outlined here provide a comprehensive approach for maintaining, expanding, and differentiating the GCaMP hiPSCs, video recording of calcium transients, and the subsequent signal extraction, preprocessing, analysis, and data visualization. © 2024 The Author(s). Current Protocols published by Wiley Periodicals LLC. Basic Protocol 1: Maintenance, expansion, and differentiation of genetically encoded calcium indicator hiPSCs Support Protocol: Replating GCaMP hiPSC-CMs for stimulation and multielectrode array studies Basic Protocol 2: Video recording from calcium transients of GCaMP hiPSC-CMs Basic Protocol 3: Signal extraction, preprocessing, analysis, and data visualization.

Magnetic 3D bioprinting of skeletal muscle spheroid for a spheroid-based screening assay

Over the past decade, there has been a rapid development in the use of magnetic three dimensional (3D) based cell culture systems. Concerning the skeletal muscle, 3D culture systems can provide biological insights for translational clinical research in the fields of muscle physiology and metabolism. These systems can enhance the cell culture environment by improving spatially-oriented cellular assemblies and morphological features closely mimicking the in vivo tissues/organs, since they promote strong interactions between cells and the extracellular matrix (ECM). However, the time-consuming and complex nature of 3D traditional culture techniques pose a challenge to the widespread adoption of 3D systems. Herein, a bench protocol is presented for creating an innovative, promptly assembled and user-friendly culture platform for the magnetic 3D bioprinting of skeletal muscle spheroids. Our protocol findings revealed consistent morphological outcomes and the functional development of skeletal muscle tissue, as evidenced by the expression of muscle-specific contractile proteins and myotubes and the responsiveness to stimulation with cholinergic neurotransmitters. This proof-of-concept protocol confirmed the future potential for further validation and application of spheroid-based assays in human skeletal muscle research.

DISCUSSING THE ISSUES OF BUILDING A SYSTEM OF TRAINING LOCAL CHINESE LANGUAGE TEACHERS IN KAZAKHSTAN

Under the framework of the Belt and Road Initiative, this study focuses on the construction and optimisation of the local Chinese language teacher education system in Kazakhstan. By examining in detail, the structure and function of the current system of Chinese language teacher training in Kazakhstan and the multiple challenges it faces, we use comparative pedagogical methods and refer to successful models of teacher education at home and abroad, with the aim of proposing a set of actionable strategic recommendations. These proposals aim not only to promote the professional growth and stability of local Chinese language teachers, but also to enhance the quality and effi-ciency of Chinese language teaching, thus promoting the popularisation and application of Chinese language in Kazakhstan. Srengthening international co-operation in the field of language education, especially exchanges with China, iscrucial for enhancing understanding and deepening the human-istic bond between the two peoples. Therefore, this study also aims to provide theoretical support and practical guidance for the construction of a shared linguistic and cultural platform connected two countries and promoted of the regional practice of the concept "community of human destiny", thus contributing to the pluralistic dialogue and harmonious coexistence among the countries along the "Belt and Road".

Robust tissue pattern formation by coupling morphogen signal and cell adhesion.

Morphogens, locally produced signaling molecules, form a concentration gradient to guide tissue patterning. Tissue patterns emerge as a collaboration between morphogen diffusion and responsive cell behaviors, but the mechanisms through which diffusing morphogens define precise spatial patterns amidst biological fluctuations remain unclear. To investigate how cells respond to diffusing proteins to generate tissue patterns, we develop SYMPLE3D, a 3D culture platform. By engineering gene expression responsive to artificial morphogens, we observe that coupling morphogen signals with cadherin-based adhesion is sufficient to convert a morphogen gradient into distinct tissue domains. Morphogen-induced cadherins gather activated cells into a single domain, removing ectopically activated cells. In addition, we reveal a switch-like induction of cadherin-mediated compaction and cell mixing, homogenizing activated cells within the morphogen gradient to form a uniformly activated domain with a sharp boundary. These findings highlight the cooperation between morphogen gradients and cell adhesion in robust tissue patterning and introduce a novel method for tissue engineering to develop new tissue domains in organoids.

Increased Susceptibility to Mechanical Stretch Drives the Persistence of Keloid Fibroblasts: An Investigation Using a Stretchable PDMS Platform.

Keloids are a common fibrotic disease of the skin, with the pathological hallmark of excessive extracellular matrix synthesis due to abnormal fibroblast activity. Since keloids clinically arise in areas of high mechanical tension, the mechanotransductory pathway may be attributed to its pathogenesis. We aimed to establish a preclinical platform to elucidate the underlying mechanism of keloid development and its clinical persistence. We fabricated a mechanically stretchable polydimethylsiloxane cell culture platform; with its mimicry of the in vivo cyclic stretch of skeletal muscles, cells showed higher proliferation compared with conventional modalities. In response to mechanical strain, TGF-β and type 1 collagen showed significant increases, suggesting possible TGF-β/Smad pathway activation via mechanical stimulation. Protein candidates selected by proteomic analysis were evaluated, indicating that key molecules involved in cell signaling and oxidative stress were significantly altered. Additionally, the cytoskeletal network of keloid fibroblasts showed increased expression of its components after periodic mechanical stimulation. Herein, we demonstrated and validated the existing body of knowledge regarding profibrotic mechanotransduction signaling pathways in keloid fibroblasts. Cyclic stretch, as a driving force, could help to decipher the tension-mediated biomechanical processes, leading to the development of optimized therapeutic targets.