-
The Journal of Physiology Feb 2022Adipocyte enlargement is a key feature of obesity and associated with insulin resistance and metabolic disease The cause and consequences of adipocyte enlargement have...
Adipocyte enlargement is a key feature of obesity and associated with insulin resistance and metabolic disease The cause and consequences of adipocyte enlargement have remained hard to study in vitro due to a lack of human cell models with representative morphology This paper provides an easily set up spheroid culture method, HUVAS (human unilocular vascularized adipocyte spheroids), for the differentiation and culturing of human adipocytes with a more unilocular morphology We show that providing adipocyte progenitors with a vascular differentiation niche is key for achieving in vitro differentiated adipocytes with large lipid droplets Lipid treatment of the HUVAS spheroids can further adipocyte enlargement and induce cellular dysfunction, mimicking the in vivo effects of weight gain The model will allow a wider research community to perform mechanistic studies of the factors impacting on human adipocyte differentiation and growth, increasing our understanding of how obesity develops and why it has such detrimental consequences on whole body metabolism ABSTRACT: The rise in obesity prevalence has created an urgent need for new and improved methods to study human adipocytes and the pathogenic effects of weight gain in vitro. Despite the proven advantage of culturing adipocyte progenitors as 3D structures, the majority of studies continue to use traditional 2D cultures which result in small, multilocular adipocytes with poor representability. We hypothesized that providing differentiating pre-adipocytes with a vascular growth niche would mimic in vivo adipogenesis and improve the differentiation into unilocular adipocytes. Here we present HUVAS (human unilocular vascularized adipocyte spheroids), a simple, easily applicable culture protocol that allows for the differentiation of human adipocytes with a more unilocular morphology and larger lipid droplets than previous protocols. Moreover, we offer a protocol for inducing adipocyte enlargement in vitro, resulting in larger lipid droplets and development of several key features of adipocyte dysfunction, including altered adipokine secretion, impaired lipolysis and insulin resistance. Taken together, our HUVAS model offers an improved culture system for studying the cellular and molecular mechanisms causing metabolic dysfunction and inflammation in human adipose tissue during weight gain.
Topics: Adipocytes; Adipogenesis; Adipose Tissue; Cell Differentiation; Humans; Weight Gain
PubMed: 34387376
DOI: 10.1113/JP281445 -
Therapeutic and prophylactic uses of invertebrates in contemporary Spanish ethnoveterinary medicine.Journal of Ethnobiology and... Sep 2016Zootherapeutic practices in ethnoveterinary medicine are important in many socio-cultural environments around the world, particularly in developing countries, and they... (Review)
Review
Zootherapeutic practices in ethnoveterinary medicine are important in many socio-cultural environments around the world, particularly in developing countries, and they have recently started to be inventoried and studied in Europe. In light of this, the purpose of this review is to describe the local knowledge and folk remedies based on the use of invertebrates and their derivative products in contemporary Spanish ethnoveterinary medicine. An overview in the fields of ethnozoology, ethnoveterinary medicine and folklore was made. Automated searches in the most important databases were performed. All related works were examined thoroughly and use-reports were obtained from 53 documentary sources. The traditional use of 18 invertebrate species and five ethnotaxa and a total of 86 empirical remedies based on the use of a single species was recorded. The two most relevant zoological groups were found to be insects and molluscs. A broad diversity of body parts or derivative products have been and are used to treat or prevent ca. 50 animal diseases or conditions, in particular diseases of the skin and subcutaneous tissue, different infectious livestock diseases, and disorders of the eye and adnexa. Cattle, sheep and equines form the group of domestic animals in which the greatest number of remedies are mentioned. In addition, seven magical remedies and practices are documented. In comparison with other culturally related areas, this is a rich heritage. The use-reports included here will help in the search for new and low-cost drugs for treating livestock and alternative materials for pharmaceutical purposes, future research addressing the validation of the effects and the development of organic farming.
Topics: Animals; Invertebrates; Medicine, Traditional; Spain; Veterinary Medicine
PubMed: 27595672
DOI: 10.1186/s13002-016-0111-1 -
International Journal of Molecular... May 2020In the last decade, the generation of cardiac disease models based on human-induced pluripotent stem cells (hiPSCs) has become of common use, providing new opportunities... (Review)
Review
In the last decade, the generation of cardiac disease models based on human-induced pluripotent stem cells (hiPSCs) has become of common use, providing new opportunities to overcome the lack of appropriate cardiac models. Although much progress has been made toward the generation of hiPSC-derived cardiomyocytes (hiPS-CMs), several lines of evidence indicate that two-dimensional (2D) cell culturing presents significant limitations, including hiPS-CMs immaturity and the absence of interaction between different cell types and the extracellular matrix. More recently, new advances in bioengineering and co-culture systems have allowed the generation of three-dimensional (3D) constructs based on hiPSC-derived cells. Within these systems, biochemical and physical stimuli influence the maturation of hiPS-CMs, which can show structural and functional properties more similar to those present in adult cardiomyocytes. In this review, we describe the latest advances in 2D- and 3D-hiPSC technology for cardiac disease mechanisms investigation, drug development, and therapeutic studies.
Topics: Cardiovascular Diseases; Cell Culture Techniques; Cell Differentiation; Cells, Cultured; Humans; Induced Pluripotent Stem Cells; Models, Cardiovascular; Myocytes, Cardiac; Tissue Engineering; Tissue Scaffolds
PubMed: 32403456
DOI: 10.3390/ijms21093404 -
Journal of Tissue Viability May 2023Soft tissue material properties are vital to human body models that evaluate interactions between the human body and its environment. Such models evaluate internal... (Review)
Review
Soft tissue material properties are vital to human body models that evaluate interactions between the human body and its environment. Such models evaluate internal stress/strain responses in soft tissues to investigate issues like pressure injuries. Numerous constitutive models and parameters have been used to represent mechanical behavior of soft tissues in biomechanical models under quasi-static loading. However, researchers reported that generic material properties cannot accurately represent specific target populations due to large inter-individual variability. Two challenges that exist are experimental mechanical characterization and constitutive modeling of biological soft tissues and personalization of constitutive parameters using non-invasive, non-destructive bedside testing methods. It is imperative to understand the scope and appropriate applications for reported material properties. Thus, the goal of this paper was to compile studies from which soft tissue material properties were obtained and categorize them by source of tissue samples, methods used to quantify deformation, and material models used to describe tissues. The collected studies displayed wide ranges of material properties, and factors that affected the properties included whether tissue samples were in vivo or ex vivo, from humans or animals, the body region tested, body position during in vivo studies, deformation measurements, and material models used to describe tissues. Because of the factors that affected reported material properties, it is clear that much progress has been made in understanding soft tissue responses to loading, yet there is a need to broaden the scope of reported soft tissue material properties and better match reported properties to appropriate human body models.
Topics: Animals; Humans; Human Body; Stress, Mechanical; Biomechanical Phenomena; Finite Element Analysis; Elasticity
PubMed: 36878737
DOI: 10.1016/j.jtv.2023.02.003 -
The British Journal of Ophthalmology Jul 2022To report the global uptake of simple limbal epithelial transplantation (SLET) and compare the economic, clinical and social outcomes of SLET with those of cultured... (Review)
Review
AIMS
To report the global uptake of simple limbal epithelial transplantation (SLET) and compare the economic, clinical and social outcomes of SLET with those of cultured limbal epithelial transplantation (CLET).
METHODS
A comprehensive literature review and an online survey of eye surgeons were conducted to understand the efficacy and current uptake of SLET surgery. A de novo economic model was developed to estimate the cost savings with SLET compared with CLET. Our economic analysis is conducted from an Indian perspective, as this is where the technique originated. A scenario analysis using the UK cost data and a user-friendly Excel model is included to allow users to input the costs from their setting to estimate the cost savings with using SLET compared with using CLET RESULTS: The anatomical success with SLET in adults (72.6% (range 62%-80%)) was the same as CLET (70.4% (range 68%-80.9%)). For children, the outcome for SLET (77.8% (range 73%-83%)) was better than with CLET (44.5% (range 43%-45%)). In response to our informal questionnaire, 99 surgeons reported to have performed SLET on 1174 patients in total. They appreciated that SLET negates the requirement for costly tissue engineering facilities. Results of economic analysis suggested that SLET provided an estimated cost-savings of US$6470.88 for adults and US$6673.10 for children. In broad terms, the cost of SLET is approximately 10% of the cost of CLET for adults and 8% for children.
CONCLUSION
SLET offers a more accessible and financially attractive alternative to CLET to treat limbal stem cell deficiency.
Topics: Adult; Child; Corneal Diseases; Epithelium, Corneal; Humans; Limbus Corneae; Scleral Diseases; Social Change; Stem Cell Transplantation; Transplantation, Autologous
PubMed: 33688000
DOI: 10.1136/bjophthalmol-2020-318642 -
Biotechnology and Bioengineering Jan 1998The engineering of functional smooth muscle (SM) tissue is critical if one hopes to successfully replace the large number of tissues containing an SM component with...
The engineering of functional smooth muscle (SM) tissue is critical if one hopes to successfully replace the large number of tissues containing an SM component with engineered equivalents. This study reports on the effects of SM cell (SMC) seeding and culture conditions on the cellularity and composition of SM tissues engineered using biodegradable matrices (5 x 5 mm, 2-mm thick) of polyglycolic acid (PGA) fibers. Cells were seeded by injecting a cell suspension into polymer matrices in tissue culture dishes (static seeding), by stirring polymer matrices and a cell suspension in spinner flasks (stirred seeding), or by agitating polymer matrices and a cell suspension in tubes with an orbital shaker (agitated seeding). The density of SMCs adherent to these matrices was a function of cell concentration in the seeding solution, but under all conditions a larger number (approximately 1 order of magnitude) and more uniform distribution of SMCs adherent to the matrices were obtained with dynamic versus static seeding methods. The dynamic seeding methods, as compared to the static method, also ultimately resulted in new tissues that had a higher cellularity, more uniform cell distribution, and greater elastin deposition. The effects of culture conditions were next studied by culturing cell-polymer constructs in a stirred bioreactor versus static culture conditions. The stirred culture of SMC-seeded polymer matrices resulted in tissues with a cell density of 6.4 +/- 0.8 x 10(8) cells/cm3 after 5 weeks, compared to 2.0 +/- 1.1 x 10(8) cells/cm3 with static culture. The elastin and collagen synthesis rates and deposition within the engineered tissues were also increased by culture in the bioreactors. The elastin content after 5-week culture in the stirred bioreactor was 24 +/- 3%, and both the elastin content and the cellularity of these tissues are comparable to those of native SM tissue. New tissues were also created in vivo when dynamically seeded polymer matrices were implanted in rats for various times. In summary, the system defined by these studies shows promise for engineering a tissue comparable in many respects to native SM. This engineered tissue may find clinical applications and provide a tool to study molecular mechanisms in vascular development.
Topics: Animals; Biocompatible Materials; Biodegradation, Environmental; Biomedical Engineering; Biotechnology; Culture Techniques; Microscopy, Electron, Scanning; Muscle Development; Muscle, Smooth, Vascular; Polymers; Rats; Time Factors
PubMed: 10099177
DOI: No ID Found -
Scientific Reports Aug 2021Organoids culture provides unique opportunities to study human diseases and to complement animal models. Several organs and tissues can be in vitro cultured in 3D...
Organoids culture provides unique opportunities to study human diseases and to complement animal models. Several organs and tissues can be in vitro cultured in 3D structures resembling in vivo tissue organization. Organoids culture contains most of the cell types of the original tissue and are maintained by growth factors mimicking the in vivo state. However, the system is yet not fully understood, and specific in vivo features especially those driven by cell-extrinsic factors may be lost in culture. Here we show a comprehensive transcriptome-wide characterization of mouse gut organoids derived from different intestinal compartments and from mice of different gender and age. RNA-seq analysis showed that the in vitro culture strongly influences the global transcriptome of the intestinal epithelial cells (~ 60% of the total variance). Several compartment-, age- and gender-related transcriptome features are lost after culturing indicating that they are driven by niche or systemic factors. However, certain intrinsic transcriptional programs, for example, some compartment-related features and a minority of gender- and aging- related features are maintained in vitro which suggested possibilities for these features to be studied in this system. Moreover, our study provides knowledge about the cell-extrinsic or cell-intrinsic origin of intestinal epithelial transcriptional programs. We anticipated that our characterization of this in vitro system is an important reference for scientists and clinicians using intestinal organoids as a research model.
Topics: Animals; Cells, Cultured; Female; Gene Expression Profiling; Intestine, Small; Male; Mice, Inbred C57BL; Organoids; Stem Cells; Tissue Culture Techniques; Transcriptome; Mice
PubMed: 34404908
DOI: 10.1038/s41598-021-96321-8 -
STAR Protocols Mar 2021Patient-derived tumor organoid cultures are an essential and innovative methodology for translational research. However, current techniques to establish these cultures...
Patient-derived tumor organoid cultures are an essential and innovative methodology for translational research. However, current techniques to establish these cultures are cumbersome, expensive, and often require irreplaceable clinical tissue from surgery or core biopsies. Fine-needle aspiration (FNA) provides a minimally invasive biopsy technique commonly performed in clinical settings. Here, we provide a protocol for FNA. We have found that FNA provides a cost-effective, rapid, and streamlined method for tissue acquisition for cancer organoid culture. For complete details on the use and execution of this protocol, please refer to Lee et al. (2020) and Vilgelm et al. (2020).
Topics: Biopsy, Fine-Needle; Female; Humans; Male; Neoplasms; Organoids; Tumor Cells, Cultured
PubMed: 33377121
DOI: 10.1016/j.xpro.2020.100220 -
Scientific Reports Sep 2023The field of wireless communication has witnessed tremendous advancements in the past few decades, leading to more pervasive and ubiquitous networks. Human bodies are...
The field of wireless communication has witnessed tremendous advancements in the past few decades, leading to more pervasive and ubiquitous networks. Human bodies are continually exposed to electromagnetic radiation, but typically this does not impact the body as the radiation is non-ionizing and the waves carry low power. However, with progress in the sixth generation (6G) of wireless networks and the adoption of the spectrum above 100 GHz in the next few years, higher power radiation is needed to cover larger areas, exposing humans to stronger and more prolonged radiation. Also, water has a high absorption coefficient at these frequencies and could lead to thermal effects on the skin. Hence, there is a need to study the radiation effects on human tissues, specifically the photothermal effects. In this paper, we present a custom-built, multi-physics model to investigate electromagnetic wave propagation in human tissue and study its subsequent photothermal effects. The proposed finite-element model consists of two segments-the first one estimates the intensity distribution along the beam path, while the second calculates the increase in temperature due to the wave distribution inside the tissue. We determine the intensity variation in the tissue using the radiative transfer equation and compare the results with Monte Carlo analysis and existing analytical models. The intensity information is then utilized to predict the rise in temperature with a bio-heat transfer module, powered by Pennes' bioheat equation. The model is parametric, and we perform a systematic photothermal analysis to recognize the crucial variables responsible for the temperature growth inside the tissue, particularly for terahertz and near-infrared optical frequencies. Our numerical model can serve as a benchmark for studying the high-frequency radiation effects on complex heterogeneous media such as human tissue.
Topics: Humans; Animals; Benchmarking; Communication; Culture; Estrus; Hot Temperature
PubMed: 37669995
DOI: 10.1038/s41598-023-41808-9 -
Journal of Animal Science Aug 2020Proof-of-principle for large-scale engineering of edible muscle tissue, in vitro, was established with the product's introduction in 2013. Subsequent research and...
Proof-of-principle for large-scale engineering of edible muscle tissue, in vitro, was established with the product's introduction in 2013. Subsequent research and commentary on the potential for cell-based meat to be a viable food option and potential alternative to conventional meat have been significant. While some of this has focused on the biology and engineering required to optimize the manufacturing process, a majority of debate has focused on cultural, environmental, and regulatory considerations. Animal scientists and others with expertise in muscle and cell biology, physiology, and meat science have contributed to the knowledge base that has made cell-based meat possible and will continue to have a role in the future of the new product. Importantly, the successful introduction of cell-based meat that looks and tastes like conventional meat at a comparable price has the potential to displace and/or complement conventional meat in the marketplace.
Topics: Animals; Consumer Behavior; Culture; Food Preferences; Food Technology; Humans; Meat; Muscle, Skeletal; Stem Cells; Tissue Culture Techniques; United States; United States Food and Drug Administration
PubMed: 32745187
DOI: 10.1093/jas/skaa177