Hasil untuk "Animal culture"

Waad H. Abuwatfa, W. Pitt, G. Husseini

Three-dimensional (3D) cell cultures have emerged as valuable tools in cancer research, offering significant advantages over traditional two-dimensional (2D) cell culture systems. In 3D cell cultures, cancer cells are grown in an environment that more closely mimics the 3D architecture and complexity of in vivo tumors. This approach has revolutionized cancer research by providing a more accurate representation of the tumor microenvironment (TME) and enabling the study of tumor behavior and response to therapies in a more physiologically relevant context. One of the key benefits of 3D cell culture in cancer research is the ability to recapitulate the complex interactions between cancer cells and their surrounding stroma. Tumors consist not only of cancer cells but also various other cell types, including stromal cells, immune cells, and blood vessels. These models bridge traditional 2D cell cultures and animal models, offering a cost-effective, scalable, and ethical alternative for preclinical research. As the field advances, 3D cell cultures are poised to play a pivotal role in understanding cancer biology and accelerating the development of effective anticancer therapies. This review article highlights the key advantages of 3D cell cultures, progress in the most common scaffold-based culturing techniques, pertinent literature on their applications in cancer research, and the ongoing challenges.

Tatsuma Yao, Yuta Asayama

Cell culture technology has spread prolifically within a century, a variety of culture media has been designed. This review goes through the history, characteristics and current issues of animal‐cell culture media.

Francisco Acuña, Gisela Soledad Gualdoni, Francisco Rivollier et al.

During development, the remodelling of fibrillar components of the uterine extracellular matrix (ECM), mediated by the matrix metalloproteinases (MMPs) and their tissue inhibitors (TIMPs), plays an essential role in embryonic survival. Previously, we observed that in the plains viscacha (<i>Lagostomus maximus</i>), only caudal implantation sites (IS) contain viable embryos, whereas embryos at cranial and middle IS die and are reabsorbed. The objective of this study was to analyse the distribution and expression of key components of the endometrial ECM, including fibrillar collagens, MMPs 2 and 9, and TIMPs 1 and 2, in three uterine segments (US) of the non-pregnant adult viscachas. In sections from three US, we observed a significant craniocaudal increase in collagen fibres (Van Gieson and Picrosirius red staining) and elastic fibres (Verhoeff-Van Gieson trichrome staining), along with the immunolabelling levels of MMP-2, MMP-9, TIMP-1, and TIMP-2 (immunohistochemistry). Zymography revealed similar gelatinolytic activity of MMP-2 in the three US but higher than the MMP-9 activity. However, MMP-9 activity in the caudal segment was significantly higher than that in the cranial and middle ones. These findings suggest that uterine ECM variations along the craniocaudal axis may contribute to uterine remodelling processes that regulate embryonic survival during gestation.

Li-Tao Che, Li-Tao Che, Ahmed H. El-Sappah et al.

IntroductionThis study investigated whether glycyrrhetinic acid (GA) can alleviate the reproductive toxicity of Zearalenone (ZEN) in replacement gilts.MethodsEighty Landrace × Yorkshire gilts were randomly assigned to four dietary groups: control (basal diet), ZEN (1 mg/kg), GA (400 mg/kg), and ZEN + GA (1 mg/kg ZEN + 400 mg/kg GA).ResultsThe onset of estrus advanced significantly in all treatment groups, with the GA and ZEN + GA groups showing the most pronounced changes. Puberty onset occurred earlier in the ZEN group and was further advanced by GA supplementation. ZEN exposure impaired uterine and ovarian development, while GA improved organ development and mitigated the abnormalities in the ZEN + GA group. Hormonal analysis revealed that ZEN reduced estradiol (E2) and luteinizing hormone (LH), whereas GA elevated all measured hormones. The ZEN + GA group showed a partial recovery in hormone levels, excluding E2. Histological examination of liver tissue in the ZEN group revealed focal hepatocellular necrosis and lymphocyte infiltration, which GA notably attenuated. ZEN upregulated 3α/3β/17β-hydroxysteroid dehydrogenase (HSD) gene expression in the liver and duodenum, while GA co-administration downregulated most HSD genes except hepatic 3α-HSD.Discussion and conclusionThese findings suggest that GA can alleviate ZEN-induced reproductive toxicity via modulation of endocrine and hepatic metabolic pathways.

Souvarish Sarkar, Emir Malovic, Dilshan S. Harishchandra et al.

The NLRP3 inflammasome signaling pathway is a major contributor to the neuroinflammatory process in the central nervous system. Oxidative stress and mitochondrial dysfunction are key pathophysiological processes of many chronic neurodegenerative diseases, including Parkinson’s disease (PD). However, the inter-relationship between mitochondrial defects and neuroinflammation is not well understood. In the present study, we show that impaired mitochondrial function can augment the NLRP3 inflammasome-driven proinflammatory cascade in microglia. Primary mouse microglia treated with the common inflammogen LPS increased NLRP3 and pro-IL-1β expression. Interestingly, exposure of LPS-primed microglial cells to the mitochondrial complex-I inhibitory pesticides rotenone and tebufenpyrad specifically potentiated the NLRP3 induction, ASC speck formation and pro-IL-1β processing to IL-1β in a dose-dependent manner, indicating that mitochondrial impairment heightened the NLRP3 inflammasome-mediated proinflammatory response in microglia. The neurotoxic pesticide-induced NLRP3 inflammasome activation was accompanied by bioenergetic defects and lysosomal dysfunction in microglia. Furthermore, the pesticides enhanced mitochondrial ROS generation in primary microglia, while amelioration of mitochondria-derived ROS by the mitochondria-targeted antioxidant mito-apocynin completely abolished IL-1β release, indicating mitochondrial ROS drives potentiation of the NLRP3 inflammasome in microglia. Exposure to conditioned media obtained from mitochondrial inhibitor-treated, LPS-primed microglial cells, but not unprimed cells, induced dopaminergic neurodegeneration in cultured primary mesencephalic and human dopaminergic neuronal cells (LUHMES). Notably, our in vivo results with chronic rotenone rodent models of PD further support the activation of proinflammatory NLRP3 inflammasome signaling due to mitochondrial dysfunction. Collectively, our results demonstrate that mitochondrial impairment in microglia can amplify NLRP3 inflammasome signaling, which augments the dopaminergic neurodegenerative process. A team of American researchers demonstrate that disruption of mitochondria in microglia contributes to inflammation and neurodegeneration. Anumantha G. Kanthasamy at Iowa State University in Ames, IA and colleagues examined the effect of pesticides known to impair mitochondrial function on proinflammatory signaling pathways in microglia, the brain’s immune cells. They found that both rotenone and tebufenpyrad specifically stimulated the NLRP3 inflammasome, a multi-protein complex implicated in neuroinflammatory processes. The pesticide-treated microglia were able to cause more damage to neuronal cells than the untreated ones, indicating that mitochondrial dysfunction in microglia augments neurodegeneration. The authors also show that in rodents chronically exposed to rotenone, which causes many of the features of Parkinson’s disease (PD), the NLRP3 inflammasome is activated. These findings contribute to better understand the mechanisms driving chronic neuroinflammation in PD.

S. Genç, M. Mendeş

ABSTRACT In this study, performances of Multiple Linear Regression and Automatic Linear Modelling are compared for different sample sizes and number of predictors. A comprehensive Monte Carlo simulation study was carried out for this purpose. Random numbers generated from multivariate normal distribution by using RNMVN function of IMSL library of Microsoft FORTRAN Developer Studio composed the material of this study. Results of the simulation study showed that the sample size and the number of predictors are the main factors that lead to produce different results. Although both methods gave very similar results especially when studied with large sample sizes (n≥100), the Automatic linear modelling is preferred for analyzing data sets due to its simplicity in analyzing data and interpreting the results, ability to present results visually and providing more detailed information especially studying large complex data sets. It will be beneficial to use the Automatic linear modelling especially in analyzing massive and complex data sets for the purposes of investigating the relationships between one continuous dependent and 10 or more predictors and determine the factors that affect the response or target variable. At the same time, it will also be possible to evaluate the effect of each predictor with a more detailed response.

Stephanie Bond, Renaud Léguillette

Abstract Background Nebulized administration of dexamethasone on cytokine regulation in horses with moderate asthma has not been investigated. Objective To investigate the changes in expression of inflammatory cytokine mRNA after nebulized administration of dexamethasone treatment of horses with moderate asthma. Animals Horses with naturally occurring moderate asthma (n = 16) and healthy control horses (n = 4). All horses were kept in a dusty environment during the study. Methods Prospective, parallel, randomized, controlled, blinded clinical trial. Blood endogenous cortisol, tracheal mucus, and bronchoalveolar lavage (BAL) were sampled before and after 13 days treatment with either nebulized administration of dexamethasone (15 mg once daily) or 0.9% saline (3 mL). Treatment groups were randomly allocated via randomization function (Microsoft Excel). Amplification of target mRNA in BAL fluid (IL‐1β, IL‐4, IL‐5, IL‐6, IL‐8, IL‐10, IL‐12, IL‐17, IL‐23, IFN‐γ, Eotaxin‐2, and TNF‐α) was achieved by qPCR, and the relative expression software tool was used to analyze BAL inflammatory cytokine mRNA. Results Horses treated with nebulized administration of dexamethasone had increased relative expression of IL‐5 (1.70‐fold), IL‐6 (1.71‐fold), IL‐17 (3.25‐fold), IL‐12 (1.66‐fold), and TNF‐α (1.94‐fold), and decreased relative expression of IL‐23 (1.76‐fold; P = .04) in samples collected on Day 14, in comparison to samples collected on Day 0 (all P < .05). Horses treated with nebulized administration of saline had no significant difference in the relative expression of any gene (all P > .05). Conclusions and Clinical Importance Nebulized administration of dexamethasone was associated with increased expression of inflammatory cytokine mRNA. There was no improvement in inflammatory airway cytology associated with either dexamethasone or saline treatment.

S. Lazic, Charlie J Clarke-Williams, M. Munafo

Biologists establish the existence of experimental effects by applying treatments or interventions to biological entities or units, such as people, animals, slice preparations, or cells. When done appropriately, independent replication of the entity-intervention pair contributes to the sample size (N) and forms the basis of statistical inference. However, sometimes the appropriate entity-intervention pair may not be obvious, and the wrong choice can make an experiment worthless. We surveyed a random sample of published animal experiments from 2011 to 2016 where interventions were applied to parents but effects examined in the offspring, as regulatory authorities have provided clear guidelines on replication with such designs. We found that only 22% of studies (95% CI = 17% to 29%) replicated the correct entity-intervention pair and thus made valid statistical inferences. Approximately half of the studies (46%, 95% CI = 38% to 53%) had pseudoreplication while 32% (95% CI = 26% to 39%) provided insufficient information to make a judgement. Pseudoreplication artificially inflates the sample size, leading to more false positive results and inflating the apparent evidence supporting a scientific claim. It is hard for science to advance when so many experiments are poorly designed and analysed. We argue that distinguishing between biological units, experimental units, and observational units clarifies where replication should occur, describe the criteria for genuine replication, and provide guidelines for designing and analysing in vitro, ex vivo, and in vivo experiments.

Takao Sasaki, D. Biro

Studies of collective intelligence in animal groups typically overlook potential improvement through learning. Although knowledge accumulation is recognized as a major advantage of group living within the framework of Cumulative Cultural Evolution (CCE), the interplay between CCE and collective intelligence has remained unexplored. Here, we use homing pigeons to investigate whether the repeated removal and replacement of individuals in experimental groups (a key method in testing for CCE) alters the groups’ solution efficiency over successive generations. Homing performance improves continuously over generations, and later-generation groups eventually outperform both solo individuals and fixed-membership groups. Homing routes are more similar in consecutive generations within the same chains than between chains, indicating cross-generational knowledge transfer. Our findings thus show that collective intelligence in animal groups can accumulate progressive modifications over time. Furthermore, our results satisfy the main criteria for CCE and suggest potential mechanisms for CCE that do not rely on complex cognition. Groups of animals tend to solve tasks better than individuals, but it is unclear whether such socially-derived knowledge accumulates over time. Sasaki and Biro demonstrate that homing pigeon flocks progressively improve the efficiency of their routes by culturally accumulating knowledge across generations.

F. Ferrara, B. Hiebl, P. Kunzmann et al.

Research on animals is essential for science and medical progress. While it is still necessary to conduct this research, it is essential to apply the highest standards in animal welfare, including animal husbandry and care. Furthermore, it is important to recognize the special relationship between research animals and the people who care for them. Caring for research animals can be extremely fulfilling and meaningful, but it also comes with challenges, particularly when caring for animals experiencing pain or distress. These challenges can lead to work-related mental stress. To get more insight into the challenges of working in animal research, we organized a panel discussion at the GV-SOLAS (German Society for Laboratory Animal Science) and IGTP (Interest Group Animal Caretakers) conference 2021 about work wellbeing. This discussion was the first of its kind in Germany. The active panel contributions included the view of an ethical philosopher, a scientist, a lecturer for laboratory animal science, an animal facility manager and an animal caretaker. They gave insights from their perspective into key factors that can affect human wellbeing in animal research. Keys ideas included stigmatization of work, tension between research aims and animal wellbeing, and the importance of supportive culture to overcome work-related strains, as well as lack of education and supportive environments to cope with emotional stress in the workplace. Overall, the discussion has shown that we must also promote human wellbeing when promoting culture of care in animal research, because there is strong relationship between culture of care and individual performance.

A. Berdahl, Albert B. Kao, A. Flack et al.

Animals often travel in groups, and their navigational decisions can be influenced by social interactions. Both theory and empirical observations suggest that such collective navigation can result in individuals improving their ability to find their way and could be one of the key benefits of sociality for these species. Here we provide an overview of the potential mechanisms underlying collective navigation and review the known, and supposed, empirical evidence for such behaviour, and highlight interesting directions for future research. We further explore how both social and collective learning during group navigation could lead to the accumulation of knowledge at the population level, resulting in the emergence of migratory culture.

Y. Ho, Haojie Lu, Zhi Feng Sherman Lim et al.

Animal cells are used in the manufacturing of complex biotherapeutic products since the 1980s. From its initial uses in biological research to its current importance in the biopharmaceutical industry, many types of culture media were developed: from serum-based media to serum-free to protein-free chemically defined media. The cultivation of animal cells economically has become the ultimate goal in the field of biomanufacturing. Serum serves as a source of amino acids, lipids, proteins and most importantly growth factors and hormones, which are essential for many cell types. However, the use of serum is unfavorable due to its high price tag, increased lot-to-lot variations and potential risk of microbial contamination. Efforts are progressively being made to replace serum with recombinant proteins such as growth factors, cytokines and hormones, as well as supplementation with lipids, vitamins, trace elements and hydrolysates. While hydrolysates are more complex, they provide a diverse source of nutrients to animal cells, with potential beneficial effects beyond the nutritional value. In this review, we discuss the use of hydrolysates in animal cell culture and briefly cover the composition of hydrolysates, mode of action and potential contaminants with some perspectives on its potential role in animal cell culture media formulations in the future.

A. Verma, M. Verma, Anchal Singh

Animal cell culture technology in today’s scenario has become indispensable in the field of life sciences, which provides a basis to study regulation, proliferation, and differentiation and to perform genetic manipulation. It requires specific technical skills to carry out successfully. This chapter describes the essential techniques of animal cell culture as well as its applications.

Younsik Jeong, W. Choi, Areumi Park et al.

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

سادات صفوی, مرتضی چاجی

آزمایش حاضر با هدف بررسی اثرات استفاده از دو منبع اوره آهسته رهش و مقایسه‌ی آنها با جیره‌های فاقد اوره یا حاوی اوره معمولی در بره‌های پرواری تغذیه شده با مواد علوفه‌ای کم کیفیت انجام شد. چهار تیمار آزمایشی شامل ۱) تیمار شاهد (فاقد اوره)، تیمار ۲) ۸/۱ درصد اوره آهسته­رهش با نام تجاری اسلوژن، تیمار ۳) ۶۹/۱ درصد اوره آهسته­رهش اپتی­ژن و تیمار ۴) ۶/۱ درصد اوره معمولی بود. از 24 راس بره نر عربی با میانگین وزن 65/۲5 کیلوگرم و سن ۵ ماه استفاده شد. در مقایسه با گروه شاهد، استفاده از منابع اوره‌ی آهسته رهش، تاثیری بر مقدار مصرف خوراک و درصد قابلیت هضم ماده خشک، الیاف نامحلول در شوینده خنثی، الیاف نامحلول در شوینده اسیدی و ماده آلی نداشت. قابلیت هضم پروتئین خام در تیمار شاهد بیشترین و در تیمار حاوی اوره معمولی کم‌ترین مقدار بود (۰۵/۰P<) و شاهد با تیمار اسلوژن تفاوت نداشت. عملکرد رشد و پروار بره‌ها در کل دوره شامل وزن نهایی، میانگین افزایش وزن روزانه، کل افزایش وزن زنده‌، ضریب تبدیل و بازده خوراک تحت تاثیر تیمارهای آزمایشی قرار نگرفت. تیمارهای آزمایشی تاثیر معنی‌داری بر غلظت نیتروژن آمونیاکی، اسیدهای چرب فرار، جمعیت پروتوزوآ، pH شکمبه و فراسنجه‌های خونی نظیر غلظت گلوکز و نیتروژن اوره‌ای نداشت. جیره‌های حاوی اسلوژن و اوره معمولی، سود مناسب‌تری نسبت به جیره شاهد داشتند. در کـل یافته‌های آزمـایش نشـان داد کـه در شرایط آزمایش حاضر، نتایج تیمارهای حاوی منابع نیتروژن غیر پروتئینی با منابع پروتئینی (جیره شاهد) قابل رقابت بود. از طرفی تفاوت آشکاری بین دو منبع حاوی اوره آهسته رهش با یکدیگر، با جیره‌ی شاهد و با اوره معمولی مشاهده نشد؛ اما اسلوژن عملکرد مناسب‌تری داشت. بنابراین، می‌توان از منابع نیتروژن غیر­پروتئینی به عنوان جایگزینی برای بخشی از منابع پروتئین جیره‌‌ی بره‌های پروار استفاده کرد تا در نهایت هزینه تولید کاهش داده شود.

Ingrid Lorenz

Abstract Research on calf health and welfare has intensified in the past decades. This is an update on a review series on calf health from birth to weaning published ten years ago. Good colostrum management is still recognised as the single most important factor to preventing calf morbidity and mortality, however, it is now known that immunoglobulins are only one of many components of colostrum that are vital for the calf’s development. Other non-nutrient factors like leucocytes, hormones and growth factors, oligosaccharides as well as microRNAs have significant effects on the development and maturation of the intestinal and systemic immune functions. They also promote the maturation and function of the intestine, thus enabling the calf to digest and absorb the nutrients provided with colostrum and milk. The improved energetic status of colostrum-fed neonates is reflected by an accelerated maturation of the somatotropic axis, which stimulates body growth and organ development. Colostrum oligosaccharides are presumed to play a major role in the development of a healthy intestinal flora. A biologically normal (intensive) milk-feeding programme is subsequently necessary for optimal body growth, organ development and resistance to infectious diseases. Ad-libitum or close to ad-libitum feeding in the first three to four weeks of life also leaves calves less hungry thus improving calf welfare. Only calves fed intensively with colostrum and milk are able to reach their full potential for performance throughout their life. Public interest in farm animal welfare is growing in past decades, which makes it necessary to have a closer look at contentious management practices in the dairy industry like early separation of the dairy calf from the dam with subsequent individual housing. Public objection to these practices cannot be mitigated through educational efforts. Contrary to common opinion there is no evidence that early cow-calf separation is beneficial for the health of calf or cow. There is evidence of behavioural and developmental harm associated with individual housing in dairy calves, social housing improves feed intake and weight gains, and health risks associated with grouping can be mitigated with appropriate management. In conclusion, there are still many management practices commonly applied, especially in the dairy industry, which are detrimental to health and welfare of calves.

Y Fakri Mustafa, R Riyadh Khalil, E Tareq Mohammed et al.

Coumarin (2H-1-benzopyran-2-one) is a plant-derived natural product known for its pharmacological properties, such as anti-inflammatory, anticoagulant, antibacterial, antifungal, antiviral, anticancer, antihypertensive, antitubercular, anticonvulsant, antiadipogenic, antihyperglycemic, antioxidant, and neuroprotective. Two coumarin-based products were identified in the seeds of two apple phenotypes commonly known as Granny Smith and Red Delicious. This study aimed to evaluate the chemical manipulation of these coumarin-based products to more lipophilic semisynthetic compounds and trace the role of the phenolic hydroxyl group in the bioactivity of the parent natural products. The bioactivity evaluation included studying the potentials of the natural- and semisynthetic-coumarins as antioxidant, antineoplastic, antifungal, and antibacterial agents. At the first step, the antiradical potential of these products was evaluated versus the free radicals of hydroxyl and DPPH. The second potential was investigated utilizing an MTT-based photo assay versus several cancer-line cells,including SK-OV-3, MCF-7, KYSE-30, LC540, HeLa, AR42J, AB12, and AMN3. The third and fourth potentials were recognized by conducting a disc-diffusion method against six infective bacterial strains and three fungal strains. The test bacteria were Shigella dysenteriae, Klebsiella pneumonia, Escherichia coli, Haemophilus influenzae, Salmonella typhi, and Pseudomonas aeruginosa. On the other hand, the test fungi included Aspergillus flavus, Candida albicans, and Aspergillus niger. The results arising from these biopotentials revealed that the investigated functional group exerted a positive impact on the antiradical and antineoplastic potentials of the natural derivatives; however, they had a negative consequence on their antimicrobial potentials.

R. Boyd

Human beings have evolved to become the most dominant species on Earth. This astonishing transformation is usually explained in terms of cognitive ability—people are just smarter than all the rest. But this book argues that culture—our ability to learn from each other—has been the essential ingredient of our remarkable success. The book shows how a unique combination of cultural adaptation and large-scale cooperation has transformed our species and assured our survival—making us the different kind of animal we are today. The book is based on the Tanner Lectures delivered at Princeton University, featuring challenging responses across the chapters.

G. Davies, B. Greenhough, P. Hobson-West et al.

The principles of the 3Rs—replacement, refinement, and reduction—strongly shape discussion of methods for performing more humane animal research and the regulation of this contested area of technoscience. This special issue looks back to the origins of the 3Rs principles through five papers that explore how it is enacted and challenged in practice and that develop critical considerations about its future. Three themes connect the papers in this special issue. These are (1) the multiplicity of roles enacted by those who use and care for animals in research, (2) the distribution of “feelings that matter” across species and spaces of laboratory animal practice, and (3) the growing importance of “cultures of care” in animal research.

Argang Kazemzadeh, C. Elias, Melih Tamer et al.

Abstract The single-use stirred bioreactors are increasingly recognized as a viable alternative in animal cell culture due to the higher production capacity, increased flexibility, prevention of cross contamination, reduction of the cleaning cost, and shortened downtime. In this paper, the effects of the impeller speed, the volumetric gas flow rate, and the impeller type on the volumetric gas-liquid mass transfer coefficient in a single-use unbaffled angled-shaft bioreactor applicable for the animal cell culture were analyzed and compared with those attained for the baffled vertical-shaft bioreactors. The volumetric gas-liquid mass transfer coefficient (KLa) was experimentally determined by the simplified dynamic pressure method. The lowest P/V values as well as the highest KLa values were achieved for an unbaffled angled-shaft bioreactor. It was found that the Rushton impeller was the most efficient impeller in term of mass transfer for this bioreactor. An empirical correlation for the mass transfer coefficient was also developed.