Culture Expansion Research Articles

Life-long functional regeneration of in vivo airway epithelium by the engraftment of airway basal stem cells.

Durable and functional regeneration of the airway epithelium in vivo with transplanted stem cells has the potential to reconstitute healthy tissue in diseased airways, such as in cystic fibrosis or primary ciliary dyskinesia. Here, we present detailed protocols for the preparation and culture expansion of murine primary and induced pluripotent stem cell-derived airway basal stem cells (iBCs) and methods for their intra-airway transplantation into polidocanol-conditioned murine recipients to achieve durable in vivo airway regeneration. Reconstitution of the airway tissue resident epithelial stem cell compartment of immunocompetent mice with syngeneic donor cells leverages the extensive self-renewal and multipotent differentiation properties of basal stem cells (BCs) to durably generate a broad diversity of mature airway epithelial lineages in vivo. Engrafted donor-derived cells re-establish planar cell polarity as well as functional ciliary transport. By using this same approach, human primary BCs or iBCs transplanted into NOD-SCID gamma recipient mice similarly display engraftment and multilineage airway epithelial differentiation in vivo. The time to generate mouse or human iBCs takes ~60 d, which can be reduced to ~20 d if previously differentiated cells are thawed from cryopreserved iBC archives. The tracheal conditioning regimen and cell transplantation procedure is completed in 1 d. A competent graduate student or postdoctoral trainee should be able to perform the procedures listed in this protocol.

Culture Expansion Alters Human Bone Marrow Derived Mesenchymal Stem Cell Production of Osteoarthritis-relevant Cytokines and Growth Factors

PurposeThe purposes of this study were to characterize the human bone marrow derived mesenchymal stem cells (BM-MSCs) production of osteoarthritis-relevant cytokines and growth factors as they are purified and multiplied, a process termed culture expansion, and to compare the immunomodulatory potential of BM-MSCs based on source and medium used for culture expansion. MethodsBM-MSCs were obtained from iliac crest bone marrow aspirates of four healthy donors. These four BM-MSC cell lines underwent four rounds, or “passages,” of the institutional culture expansion protocol, using institutional culture media. The secretory molecules known to play a role in OA-related inflammatory immune response, cartilage degradation and patient symptoms, together called the BM-MSC “secretome,” were measured at each passage. Three lines of commercially available BM-MSCs from healthy donors underwent culture expansion by the same protocol, using commercial culture media. The commercial BM-MSCs secretome and the institutional BM-MSCs secretome were compared at each passage. Significance was set at p<0.05. ResultsInstitutional BM-MSCs produced less IL-6 at passages 3 (237±113 pg/mL) and 4 (237±113 pg/mL) compared to passages 1 (884±97 pg/mL) and 2 (1071±129 pg/mL; p<0.01). Institutional BM-MSCs produced more MIP-3α at passage 4 than at passage 1 (106±41 vs 32±7 pg/mL; p<0.01). Across passages of culture expansion, institutional BM-MSCs grown on institutional medium expressed more IL-6 (P<0.001), IL-10 (P<0.001), IL-1β (P<0.001), TNFα (P=0.004), and VEGF-C (P=0.003) than commercially available BM-MSCs grown on commercial medium. ConclusionCulture expansion alters key molecules within the BM-MSC secretome. Additionally, differences in BM-MSC source and culture medium alter the BM-MSC secretome and its immunomodulatory potential.

Ambiguous AI: Lossy Compression, Model Collapse, and Intercultural Sensitivity

This paper examines the intersections of artificial intelligence (AI), cultural representation, and intercultural sensitivity, critiquing the impact of lossy compression and model collapse on cultural expression in AI systems. The study introduces “intentional patiency” as an alternative framework, conceptualizing AI not merely as an active agent but as a cultural receptor, and in turn, producer. AI systems often rely on compression algorithms that strip away cultural complexities, rendering nuanced traditions as simplified, stereotypical artifacts. Drawing on frameworks from Ted Chiang’s metaphor of lossy compression to Wendy Chun’s critique of noise, the essay explores how AI’s data optimization selectively filters cultural diversity, perpetuating dominant narratives while silencing marginalized voices. Model collapse in generative systems demonstrates while also exacertabting the pitfalls of homogenous data to reduce cultural representations to monolithic “single stories,” echoing concerns raised by Chimamanda Ngozi Adichie. To address this issue, this essay advocates for an interculturally sensitive AI development paradigm, incorporating cultural expansion and ethical safeguards to mitigate cultural erasure. Through the concept of “ambiguous AI,” a paradigm rooted in probabilistic engagement and intentional patiency, the essay seeks to reframe AI’s role as a nuanced participant in cultural discourse, promoting culturally adaptive architectures.

Effects of Clump Size on the Pluripotency and Proliferation in the Passaging Process of Mouse Induced Pluripotent Stem Cells

Induced pluripotent stem cells (iPSCs) are a promising cell source because of their pluripotency and self-renewal abilities. However, there is a risk of pluripotency loss during cell expansion. Particularly, cell passaging is associated with a higher risk of decreasing cell quality. There are two iPSC passaging methods: single-cell and clump passaging. Single-cell passaging is a rapid and simple method for cell manipulation, whereas clump passaging is superior for maintaining iPSC pluripotency. Therefore, clump passaging is a robust method for expanding iPSCs while maintaining their pluripotency. However, clump size control during clump passaging is difficult because colony fragmentation is performed manually by pipetting the colonies detached from the cell culture substrates. In this study, the effect of pipetting on iPSC colony fragmentation was evaluated and the relationship between iPSC clump size and pluripotency was clarified. An automated pipetting device was developed to standardize the clump passage process. The effect of clump size on the pluripotency and proliferative capacity of mouse iPSCs was investigated. Clump size was controlled by varying the number of pipetting cycles, and pluripotency and proliferation were assessed via alkaline phosphatase staining and flow cytometry. Our results revealed that a decrease in clump size corresponded to an increase in cell proliferation, while pluripotency maintenance was optimized under specific clump sizes. These results underscore the significance of clump size for stem cell quality, emphasizing the need for a balanced approach to maintain pluripotency while fostering proliferation in the cell expansion culture for iPSCs.

GLOBAL IDENTITY AS A TYPE OF SOCIAL IDENTITY

Globalization has affected, perhaps more than any other force, the way people live, configure their physical environment, create the institutions that govern their activities, and build the markets in which they exchange resources, goods, and services. Economic and cultural exchanges brought about by waves of globalization have led to changes in consumer demographics and psychographics, i.e. changes in consumer aspirations, motivations, and behaviour. Advances in technology, transportation, global exports, and media innovation have made global flows of information, products, and services more widespread and rapid. Cultural exchanges not only contribute to the formation of an expansive culture but also affect the psychological and social identity of people. Globalization promotes the free exchange of ideas and communication throughout the world, which leads to an increase in people’s awareness of the world around them as a whole. Globalization leads to the formation of the general consciousness of people about humanity. In its purest form, this common consciousness of people about humanity is called «global identity». The article aims to consider global identity as a type of social identity. Research methods applied: analysis and generalization of scientific literature to clarify approaches to defining the essence of global and local identity; theoretical generalization for formulating conclusions about global identity as a type of social identity. The psychological effects of local culture on identity are just as effective as the effects of global culture on identity. Local identity can take many forms, from ethnicity to nationality to even smaller local identifications such as neighborhood.

China as data coloniser? rethinking cultural production, cultural mediation, and consumer agency on Kenyan and Chinese e-commerce platforms

Has China become a neo-colonizer, exporting its cultural and economic power to the world based on its agenda of building soft power? Existing scholarship on neocolonialism and data colonialism largely focuses on how China's infrastructural expansion and increasingly platformised cultural sectors can achieve its ambitious platformised cultural sectors overseas. Yet, how China's cultural power is manifested, negotiated, or resisted in people's daily lives in a South–South setting remains under-researched and under-theorised. This article uses everyday fashion in Kenya as a case study to investigate China's cultural and economic power expansion in the Global South. We examined how cultural differences are negotiated and mediated on two Chinese(-invested) e-commerce platforms. Through focus groups and platform walkthrough method, our findings serve to enrich existing theories of cultural production–platform relationships applicable in the study of various cultural and creative sectors, to offer new understandings of how symbolic, sociopolitical and cultural meanings of fashion are constructed through divergent platform affordances, and to reveal the various forms of cultural negotiations and resistance in different contexts, multiplying our frames of reference.

Spatiotemporal topology of religious spread in a multi‐religious metropolis: A historical religious profile of Taipei City in Taiwan from 1660 to 2020

AbstractReligious identities play an important role in shaping migrants' experiences. Understanding the spatiotemporal distribution of religions enhances our knowledge of both religious expansion and religious cultures. This paper aims to leverage spatiotemporal analysis to characterise the topology of the spread of multiple religions in the Taipei metropolitan area from 1660 to 2020, including Folk Taoism (853 temples/altars, 55.39%), Christianity (306 churches, 19.87%) and Buddhism (332 temples, 21.56%). Three major factors affecting religious distribution are considered: the year of establishment of religious units (churches, temples or altars), the reclamation year of religious locations, and the economic index of religious sites. The results show that Folk Taoism and Buddhism have a long history of development in early reclamation regions with a hopping expansion pattern. However, Christianity expanded later in the later reclaimed and early reclaimed areas. The Christian religious units have highly concentrated spatial patterns and are located in the higher‐income districts, centralised in the core regions of the Taipei metropolis. Modern forms of highly institutionalised religion, such as Buddhism and Christianity, are more prevalent in the relatively higher‐income, later‐cultivated areas, which provide richer social resources for development. Folk Taoism, as a traditional religion, is maintained and developed by local communities. This macroscopic analysis helps to (1) identify the exact chronology of different growth waves and explain them in a historical context, and (2) elucidate the spatial distribution differences between institutionalised religions and folk religion. Our research shows the evolution of religious landscapes from an undeveloped city to a developed city, offering valuable insights for geographical and religious studies, and enhancing understanding of diffused and institutionalised religious expansion.

Immortalized mesenchymal stromal cells overexpressing alpha-1 antitrypsin protect acinar cells from apoptotic and ferroptotic cell death.

Chronic pancreatitis (CP) is a progressive inflammatory disorder that impairs endocrine and exocrine function. Our previous work showed that mesenchymal stem/stromal cells (MSCs) and MSCs overexpressing alpha-1 antitrypsin (AAT-MSCs) could be therapeutic tools for CP. However, primary MSCs are predisposed to undergo senescence during culture expansion, which limits their therapeutic applications. We generated and characterized immortalized human MSCs (iMSCs) and AAT-MSCs (iAAT-MSCs) and tested their protective effect on 2,4,6-Trinitrobenzenesulfonic acid (TNBS)-induced acinar cell death in an invitro cell culture system. Primary MSCs were immortalized by transduction with simian virus 40 large T antigen (SV40LT), and the resulting iMSC and iAAT-MSC lines were analysed for proliferation, senescence, phenotype and multi-differentiation potential. Subsequently, apoptosis and ferroptosis pathways were investigated by assessing changes before and after TNBS treatment. Coculture of iMSCs and iAAT-MSCs with acinar cell lines inhibited early cell death induced by TNBS, reduced ER stress and reversed TNBS-induced protein reduction at tight junctions. Additionally, iMSCs and iAAT-MSCs exerted such protection by regulating mitochondrial respiration, ATP content and ROS production in TNBS-induced acinar cells. Furthermore, iMSCs and iAAT-MSCs ameliorated TNBS-induced ferroptosis by modulating iron generation and ROS production and regulating the ferritin heavy chain 1 (FTH1)/protein disulfide isomerase (PDI)/glutathione peroxide 4 (GPX4) signalling pathways in acinar cells. These findings identify ferroptosis as an unrecognized mechanism that leads to TNBS-induced cell death and offer mechanistic insights relevant to using stem cell therapy to treat acinar cell death associated with CP.

Engineered autologous nasal cartilage for repair of nasal septal perforations - a case series.

This phase I clinical trial assessed the use of autologous nasal chondrocyte tissue-engineered cartilage (N-TEC) for functional repair of nasal septal perforations (NSP). The most widely used technique to treat NSP, namely interposition grafting with a polydioxanone (PDS) plate combined with a deep temporal fascia (DTF) graft, is still suboptimal towards patient satisfaction and revision rates. Patients (n=5, all female, age range: 23-54 years) had a 0.5-2.0cm diameter NSP. N-TEC was manufactured by expansion and 3D culture of autologous nasal septum chondrocytes into Chondro-Gide® collagen membranes. N-TEC was then shaped intraoperatively and enveloped in the harvested DTF before suturing it into the NSP. Safety (primary outcome) was assessed by the number of serious adverse reactions (SAR) until 12 months. Secondary outcomes included feasibility, assessed by surgical graft manipulation, and efficacy, assessed using subjective scoring (Nasal Obstruction Symptom Evaluation, NOSE, and Visual Analogue Scale, VAS, scores) and objective breathing function tests. Structural closure of NSP after 12 months was defined using endoscopy and computed tomography (CT) scans. NSP treatment by N-TEC implantation was safe and feasible, as no SAR and no challenge in graft manipulation was recorded for any of the patients. One year postoperative, subjective scoring improved in all patients, unless already optimal (average improvement of 23 and 28.6 points out of 100 respectively for NOSE and VAS scores). Objective respiratory function overall confirmed - with the exception of one case - the observations above (average improvement of 172 ml/s). NSP were closed and the mucosae completely healed in three patients. Autologous N-TEC is a valid treatment for NSP and warrants further clinical tests.

Phase 3 Clinical Trial Candidate Rilparencel Demonstrates Consistent Phenotypic Characteristics during Ex Vivo Culture Expansion Process

Phase 3 Clinical Trial Candidate Rilparencel Demonstrates Consistent Phenotypic Characteristics during Ex Vivo Culture Expansion Process

Further biological characterization of small molecules UM171 and SR1: In vitro effects on three hematopoietic cell populations from human cord blood

Small molecules UM171 and SR1 have already been taken into clinically-oriented protocols for the ex vivo expansion of hematopoietic stem (HSCs) and progenitor (HPCs) cells. In order to gain further insight into their biology, in the present study we have assessed their effects, both individually and in combination, on the in vitro long-term proliferation and expansion of HSCs and HPCs contained within three different cord blood-derived cell populations: MNCs (CD34+ cells = 0.8 %), LIN− cells (CD34+ cells = 41 %), and CD34+ cells (CD34+ cells >98 %). Our results show that when added to cultures in the absence of recombinant stimulatory cytokines, neither molecule had any effect. In contrast, when added in the presence of hematopoietic cytokines, UM171 and SR1 had significant stimulatory effects on cell proliferation and expansion in cultures of LIN− and CD34+ cells. No significant effects were observed in cultures of MNCs. The effects of both molecules were more pronounced in cultures with the highest proportion of CD34+ cells, and the greatest effects were observed when both molecules were added in combination. In the absence of small molecules, cell numbers reached a peak by days 25–30, and then declined; whereas in the presence of UM171 or/and SR1 cell numbers were sustained up to day 45 of culture. Our results indicate that besides CD34+ cells, LIN− cells could also be used as input cells in clinically-oriented expansion protocols, and that using both molecules simultaneously would be a better approach than using only one of them.

Metabolic modulation to improve MSC expansion and therapeutic potential for articular cartilage repair

BackgroundArticular cartilage degeneration can result from injury, age, or arthritis, causing significant joint pain and disability without surgical intervention. Currently, the only FDA cell-based therapy for articular cartilage injury is Autologous Chondrocyte Implantation (ACI); however, this procedure is costly, time-intensive, and requires multiple treatments. Mesenchymal stromal cells (MSCs) are an attractive alternative autologous therapy due to their availability and ability to robustly differentiate into chondrocytes for transplantation with good safety profiles. However, treatment outcomes are variable due to donor-to-donor variability as well as intrapopulation heterogeneity and unstandardized MSC manufacturing protocols. Process improvements that reduce cell heterogeneity while increasing donor cell numbers with improved chondrogenic potential during expansion culture are needed to realize the full potential of MSC therapy.MethodsIn this study, we investigated the potential of MSC metabolic modulation during expansion to enhance their chondrogenic commitment by varying the nutrient composition, including glucose, pyruvate, glutamine, and ascorbic acid in culture media. We tested the effect of metabolic modulation in short-term (one passage) and long-term (up to seven passages). We measured metabolic state, cell size, population doubling time, and senescence and employed novel tools including micro-magnetic resonance relaxometry (µMRR) relaxation time (T2) to characterize the effects of AA on improved MSC expansion and chondrogenic potential.ResultsOur data show that the addition of 1 mM L-ascorbic acid-2-phosphate (AA) to cultures for one passage during MSC expansion prior to initiation of differentiation improves chondrogenic differentiation. We further demonstrate that AA treatment reduced the proportion of senescent cells and cell heterogeneity also allowing for long-term expansion that led to a > 300-fold increase in yield of MSCs with enhanced chondrogenic potential compared to untreated cells. AA-treated MSCs with improved chondrogenic potential showed a robust shift in metabolic profile to OXPHOS and higher µMRR T2 values, identifying critical quality attributes that could be implemented in MSC manufacturing for articular cartilage repair.ConclusionsOur results suggest an improved MSC manufacturing process that can enhance chondrogenic potential by targeting MSC metabolism and integrating process analytic tools during expansion.

An efficient serum-free medium for ex vivo expansion of myoblasts from Larimichthys crocea for cultured meat production

The large-scale production of cultured meat has been restricted by high cost of serum, batch difference and unclear composition, thus it is important to develop serum-free media for cell growth. Here we obtained a serum-free medium (SFM), which was suitable for ex vivo culturing of myoblasts from larimichthys crocea. It offered simple composition and high effectiveness that supporting myoblast attachment, expansion and differentiation. Our findings revealed that the optimal SFM consisted of DMEM/F12 base medium, 5 mg/mL bovine serum albumin (BSA), 20 mmol HEPES, 1 % penicillin/streptomycin mixture (PS) and the exogenous additive hydrocortisone. The living cells of SFM could occupy about 65 % of the serum-containing medium (SM). Additionally, differentiation assays showed that myoblasts cultured without serum still differentiated into more muscle tubes, with a higher fusion index than those cultured with serum. Finally, transcriptome sequencing was conducted to demonstrate that hydrocortisone might act as a ligand binding to G protein-coupled receptors, triggering the downstream PI3K/AKT signaling pathway and leading to cell proliferation. Our findings will provide a basis for large-scale cell expansion and cell culture meat production in the future.

Reassessing Long-Term Cryopreservation Strategies for Improved Quality, Safety, and Clinical Use of Allogeneic Dermal Progenitor Cells

In regenerative medicine, ongoing advancements in cell culture techniques, including isolation, expansion, banking, and transport, are crucial for clinical success. Cryopreservation ensures off-the-freezer availability of living cells, enabling long-term storage and transport. Customizing cryopreservation techniques and cryoprotective agents (CPAs) for specific cell types is crucial for cell source quality, sustainability, safety, and therapeutic intervention efficiency. As regenerative medicine progresses, it becomes imperative that the scientific community and industry provide a comprehensive, cell-specific landscape of available and effective cryopreservation techniques, preventing trial-and-error approaches and unlocking the full potential of cell-based therapies. Open-sharing data could lead to safer, more efficient cell therapies and treatments. Two decades of dermal progenitor cell use for burn wound treatment and Good Manufacturing Practice-compliant technology transfers have highlighted the need for further cryopreservation optimization in manufacturing workflows. In this paper, we present experimental data assessing 5 different cryopreservation formulae for long-term storage of clinical-grade FE002 primary progenitor fibroblasts, emphasizing the crucial difference between DMSO-based and DMSO-free CPAs. Our findings suggest that CryoOx, a DMSO-free CPA, is a promising alternative yielding cell viability similar to that of established commercial CPAs. This research highlights the importance of secure, robust, and efficient cryopreservation techniques in cell banking for maximizing quality, ensuring patient safety, and advancing regenerative medicine.

Next-generation BCMA-targeted chimeric antigen receptor CARTemis-1: the impact of manufacturing procedure on CAR T-cell features.

CAR therapy targeting BCMA is under investigation as treatment for multiple myeloma. However, given the lack of plateau in most studies, pursuing more effective alternatives is imperative. We present the preclinical and clinical validation of a new optimized anti-BCMA CAR (CARTemis-1). In addition, we explored how the manufacturing process could impact CAR-T cell product quality and fitness. CARTemis-1 optimizations were evaluated at the preclinical level both, in vitro and in vivo. CARTemis-1 generation was validated under GMP conditions, studying the dynamics of the immunophenotype from leukapheresis to final product. Here, we studied the impact of the manufacturing process on CAR-T cells to define optimal cell culture protocol and expansion time to increase product fitness. Two different versions of CARTemis-1 with different spacers were compared. The longer version showed increased cytotoxicity. The incorporation of the safety-gene EGFRt into the CARTemis-1 structure can be used as a monitoring marker. CARTemis-1 showed no inhibition by soluble BCMA and presents potent antitumor effects both in vitro and in vivo. Expansion with IL-2 or IL-7/IL-15 was compared, revealing greater proliferation, less differentiation, and less exhaustion with IL-7/IL-15. Three consecutive batches of CARTemis-1 were produced under GMP guidelines meeting all the required specifications. CARTemis-1 cells manufactured under GMP conditions showed increased memory subpopulations, reduced exhaustion markers and selective antitumor efficacy against MM cell lines and primary myeloma cells. The optimal release time points for obtaining the best fit product were > 6 and < 10 days (days 8-10). CARTemis-1 has been rationally designed to increase antitumor efficacy, overcome sBCMA inhibition, and incorporate the expression of a safety-gene. The generation of CARTemis-1 was successfully validated under GMP standards. A phase I/II clinical trial for patients with multiple myeloma will be conducted (EuCT number 2022-503063-15-00).

The Dissemination, Impact and Effectiveness of Jameson’s Cultural Theory: A Survey-Based Study in the Chinese Context

This study explores the dissemination, impact, and effectiveness of Fredric Jameson’s cultural theory within Chinese mass culture by conducting a comprehensive survey across various social groups and regions. The paper examines the acceptance and communication of Jameson’s theories, focusing on how his insights into cultural expansion and ideology resonate with different demographics in China. The results indicate that the primary audience for Jameson’s works consists of individuals aged 25-35 with high educational backgrounds, with significant interest in regions like Zhejiang, Hubei, Guangzhou, Beijing, Shanghai, and Shenzhen. The survey also reveals varying degrees of familiarity with Jameson’s theories among respondents, highlighting a growing influence yet uneven understanding. These findings suggest that a systematic interpretation of Jameson’s cultural theory can significantly contribute to the development and enrichment of Chinese mass culture.

The Myofibroblast Fate of Therapeutic Mesenchymal Stromal Cells: Regeneration, Repair, or Despair?

Mesenchymal stromal cells (MSCs) can be isolated from various tissues of healthy or patient donors to be retransplanted in cell therapies. Because the number of MSCs obtained from biopsies is typically too low for direct clinical application, MSC expansion in cell culture is required. However, ex vivo amplification often reduces the desired MSC regenerative potential and enhances undesired traits, such as activation into fibrogenic myofibroblasts. Transiently activated myofibroblasts restore tissue integrity after organ injury by producing and contracting extracellular matrix into scar tissue. In contrast, persistent myofibroblasts cause excessive scarring-called fibrosis-that destroys organ function. In this review, we focus on the relevance and molecular mechanisms of myofibroblast activation upon contact with stiff cell culture plastic or recipient scar tissue, such as hypertrophic scars of large skin burns. We discuss cell mechanoperception mechanisms such as integrins and stretch-activated channels, mechanotransduction through the contractile actin cytoskeleton, and conversion of mechanical signals into transcriptional programs via mechanosensitive co-transcription factors, such as YAP, TAZ, and MRTF. We further elaborate how prolonged mechanical stress can create persistent myofibroblast memory by direct mechanotransduction to the nucleus that can evoke lasting epigenetic modifications at the DNA level, such as histone methylation and acetylation. We conclude by projecting how cell culture mechanics can be modulated to generate MSCs, which epigenetically protected against myofibroblast activation and transport desired regeneration potential to the recipient tissue environment in clinical therapies.

Strategy to Strengthen Halal Tourism

Java Province has many natural tourist attractions, especially in Trenggalek Regency. In Indonesia, the development of tourism with a halal concept is becoming a trend. Several tourist attractions are competing to create or change their tourist attractions into halal tourism so that Muslim tourists who come to visit feel safer. This study aimed to determine the critical aspects of developing halal tourism in Trenggalek Regency. This research method is qualitative, using SWOT analysis. The results of this study indicate that the potential for natural beauty in Trenggalek Regency is an aspect that can positively impact the strengthening of halal tourism and increase regional income because of the many existing opportunities. Strengthening halal tourism in Trenggalek Regency is in quadrant 1, namely in the strengths and weaknesses (SO) strategy, so that strategies to increase visitor attraction can be carried out by improving the quality of tourism by utilizing existing opportunities. Overall, this study provides a strategic framework for the development of halal tourism in Trenggalek Regency, which has broad implications for economic growth, cultural expansion, infrastructure development, stakeholder collaboration, and policy planning.

Robust bioprocess design and evaluation of commercial media for the serial expansion of human induced pluripotent stem cell aggregate cultures in vertical-wheel bioreactors

BackgroundWhile pluripotent stem cell (PSC) therapies move toward clinical and commercial applications at a rapid rate, manufacturing reproducibility and robustness are notable bottlenecks in regulatory approval. Therapeutic applications of PSCs require large cell quantities to be generated under highly robust, well-defined, and economically viable conditions. Small-scale and short-term process optimization, however, is often performed in a linear fashion that does not account for time needed to verify the bioprocess protocols and analysis methods used. Design of a reproducible and robust bioprocess should be dynamic and include a continuous effort to understand how the process will respond over time and to different stresses before transitioning into large-scale production where stresses will be amplified.MethodsThis study utilizes a baseline protocol, developed for the short-term culture of PSC aggregates in Vertical-Wheel® bioreactors, to evaluate key process attributes through long-term (serial passage) suspension culture. This was done to access overall process robustness when performed with various commercially available media and cell lines. Process output variables including growth kinetics, aggregate morphology, harvest efficiency, genomic stability, and functional pluripotency were assessed through short and long-term culture.ResultsThe robust nature of the expansion protocol was demonstrated over a six-day culture period where spherical aggregate formation and expansion were observed with high-fold expansions for all five commercial media tested. Profound differences in cell growth and quality were revealed only through long-term serial expansion and in-vessel dissociation operations. Some commercial media formulations tested demonstrated maintenance of cell growth rates, aggregate morphology, and high harvest recovery efficiencies through three bioreactor serial passages using multiple PSC lines. Exceptional bioprocess robustness was even demonstrated with sustained growth and quality maintenance over 10 serial bioreactor passages. However, some commercial media tested proved less equipped for serial passage cultures in bioreactors as cultures led to cell lysis during dissociation, reduction in growth rates, and a loss of aggregate morphology.ConclusionsThis study demonstrates the importance of systematic selection and testing of bioprocess input variables, with multiple bioprocess output variables through serial passages to create a truly reproducible and robust protocol for clinical and commercial PSC production using scalable bioreactor systems.

Analyzing the Successful Reasons on Perfect Film and Television

Due to economic growth, the cultural industry's rapid expansion, and advancements in science and technology, Perfect Word, a game company, utilized financial leverage to establish Perfect Film and Television and enter the film and television industry. This move resulted in significant profits because their film and television productions gained immense popularity among viewers. The goal of this study was to investigate the factors that contributed to Perfect World's achievements in the film and television industries. This paper examines Perfect Film and Television's precise measures, using it as a case study, and evaluates its financial statistics and the accolades it has received. The primary factor contributing to its success lies in the company's foundation of producing top-notch content, consistently broadening its distribution platforms, and venturing into international markets. To achieve further growth, Perfect Film and Television can strategically establish a comprehensive industrial chain, leverage digital technologies to broaden distribution channels, and establish a distinct traditional cultural brand. The factors that contributed to this study's success can serve as a blueprint for the expansion of other film and television enterprises.