Search Results for '3d cell culture systems'

  1. Combining High-throughput screening and 3D bioprinting

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    The future of drug screening, disease modeling, and precision medicine.

    In the last few decades, both high-throughput screening and three-dimensional (3D) bioprinting technologies have emerged and have been proven to be instrumental for the advancement of drug screening, drug development, and tissue modelling.

    High-throughput screening (HTS) is well established within the pharmaceutical industry for compound and drug discovery. It is, for the most part, the primary method of testing prior to compound validation and preclinical animal model studies prior to clinical trials. Today, the industry standard is still the use of two-dimensional (2D) cell culture for screening, as it has a long record of use and thus well-characterised cell lines and methods. This is despite the fact that the use of 2D culture is limiting due to its inability to capture in vivo-like cell-cell and cell-matrix interactions, and the fact many cell types display different phenotypes and varying genomic profiles in 2D versus 3DWhilst the advantages of 3D cell culture are well understood, the reality is that many 3d culture systems can be challenging to reproduce to the levels required for HTS.

    This is where 3D bioprintingis becoming recognised as a promising technology for HTS. It has the potential to produce more realistic, highly reproducible 3D cell cultures and support the bio-fabrication of 3D tissues and organoids.

    The future of bioprinting in pharmaceutical applications

    Bioprinting technology has advanced significantly in recent years, although it is still limited by the speed of printing and resolution. Therefore depending on the complexity of the printed construct, it may currently be a better candidate for use in lower throughput, larger scale models in 96-well or fewer plates. Nonetheless, in the future, with the advances being made in the precision, accuracy, and scale of the bioprinters, this technology might offer solutions for many current challenges of the drug development such as disease modelling, drug optimisation, toxicity screening and precision medicine.

    • Disease modelling and drug optimization

    Better disease models are crucial for better understanding the behaviour, development, treatment, prevention, and cure of disease. Bioprinting of 3D cell-hydrogel constructs, when combine with microfluidic systems, has allowed the creation of complex, reproducible organ-on-a-chip models which can replicate body wide diseases by incorporating many external features such as fluid or air flow and a combination of tissue types into a single 3d model. Such technology enables researchers to conduct disease progression studies for better understanding and ultimately treating the disease.

    • Drug toxicity screening

    When developing new drug treatments, toxic effects must be well characterised and monitored in vivo in animal models. The opportunity to more effectively model such effects prior to animal studies by utilising 3D cell culture models generated using bioprinting technology may allow for the drug to be modified or withdrawn prior to costly to in-vivo modelling.

    One of the most heavily investigated side effects in drug development is the liver toxicity since it can result from over-exposure to toxins, leading to toxic hepatitis, inflammation of the liver, and eventual cirrhosis if exposed over a long period of time. These side effects further lead to what is known as drug-induced liver injury which is life-threatening. The complexity of the liver combined with the substantial role it plays in drug metabolism has created a demand for physiologically improved liver models which can be addressed by novel bioprinting approaches and improved liver models.

    • Precision medicine

    Precision medicine is the tailoring of therapy based on a patient’s genetic information. The use of a patient’s own cells allows for an amount of data to be collected for long term studies of disease progression and drug treatment response.

    The use of 3D cell culture models in precision medicine allows for cells from the patient to be cultured and expanded to study the disease ex vivo while still maintaining in vivo genotype. Using the patient’s own cells, 3D bioprinting can be leveraged to create a large quantity of patient-specific disease models for use in parallel with clinical trials or independently through precision medicine initiatives for targeting patient or disease-specific genetic, proteomic, and phenotypic characteristics.

    In summary, the potential of 3d bioprinting applications in high-throughput screening is vast, with drug screening and toxicity studies, disease modelling, and most recently precision medicine applications having a greater potential in the future due to the increased complexity, physiological relevance and reproducibility bioprinting can offer. By using bioprinting as a tool in high-throughput screening, physiologically relevant models can be realized for the improvement of healthcare.

     

    Source:

    Andrea Mazzocchi, Shay Soker, and Aleksander Skardal (2019) Applied Physics Reviews 6, 011302; https://doi.org/10.1063/1.5056188

     

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  2. Application of Biogelx hydrogels in 3d osteoarthritis in-vitro model

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    Interview with David Hughes

    David Hughes is one of our new Biogelx sponsored PhD students.His MRS funded project aims to develop a reproducible and easily manipulated 3D osteoarthritis in vitro model by using BiogelxTM peptide hydrogelsto examine bone and cartilage crosstalk in healthy and pathological ageing. In this interview, he speaks about his research and shares his hands-on experiences working with our products.

    1. Please tell us a little bit about yourself.

    I’m a PhD student from central Scotland with a keen interest in biomedical science; I originally undertook an undergraduate degree in Biology and Psychology. Due to my interest particularly in the biological side of my studies I then went on to pursue an MSc in Biomedical Science and Drug Design. This masters gave me an insight into tissue culture and its potential research value which lead me to where I am today. Outside of my studies I am a keen guitarist and play as part of a band.

    1. What research do you focus on?

    My research is focused on the development of a novel 3D model to investigate the crosstalk between bone and cartilage in osteoarthritis (OA). Currently there is no validated in vitromodel of the whole joint for OA research and the limitations posed by current 2D techniques necessitates the development of an appropriate 3D model that is more like the in vivoconditions. My PhD is funded by Medical Research Scotland in the lab of Dr Katherine Staines at Edinburgh Napier University.

    1. Why is it so important to use synthetic 3D cell culture materials in your research?

     Synthetic 3D culture materials such as those produced by Biogelx, provide a reliable and reproducible platform that allows me to conduct a variety of techniques on my cells of interest. Importantly synthetic 3D culture systems help eliminate further factors introduced through the variability associated with the use of a biologic system.

    1. Have you tried any alternative materials? What are the key features of a good 3D cell culture?

    I have not tried any additional 3D culture materials; however, I believe that the key features of a good 3D cell culture system are biocompatibility, modifiability and ease of handling. These features can allow for the investigation of a broad spectrum of variables across a variety of cell types whilst allowing the researcher additional optimization options which can save time and provide financial benefits.

    1. How easy is to use Biogelx products?

    Biogelx products are very easy to use, it’s as simple as weighing a powder out, rehydrating it and then adding the cell culture media that contains divalent cations to promote cross-linkage. The resources required to do this are commonplace in biomedical laboratories, keeping the gel preparation process simple and accessible.

    1. What cell types do you use in your research? Please share your experience in growing cells in our gel(s). If you have any pictures that you can share, please do.

    My research is still in early stages – I’ve been working with chondrocytes in the gels and have had no issues so far. Chondrocytes are the cells found in the cartilage, which covers the ends of the bones in the joint. Preliminary imaging shows that the cells seeded on the gels take on the characteristic rounded chondrocyte shape associated with their in vivomorphology; chondrocytes plated in a monolayer do not display this and take on a fibroblast-like shape, which is not ideal for investigating these cells and their functions in vitro.

    1. What are the next steps in your research?

    The next steps in my research will involve characterising the morphology and molecular phenotype of the chondrocytes and comparing it between cells cultured in monolayers (2D) and in Biogelx hydrogels (3D). Following on from this I intend to conduct similar optimisation on cells from the other joint tissues, before looking to develop my 3D model which I will use to investigate the crosstalk between bone and cartilage in OA.

     

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  3. Why is the application of 3D Bioprinting important in Drug Development?

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    Drug development is a time-consuming and expensive process that proceeds through several stages from target identification to lead discovery and optimization, preclinical validation, and clinical trials, ending up in approval for clinical use. It is well known that 90 percent of drugs that reach clinical stage development never make it to FDA approval and commercialization. The cost of a failed drug is between $800m and £1.4bn. With this low success rate in clinical trials, drug discovery remains a slow and costly business. Hence, there is an urgent need for new technologies which can mitigate the risk of failures.

    3D bioprinting is one of the most promising areas expected to improve the success rates in drug development. Whilst most people start to imagine printed human organs for transplanting into the patients when they hear the word bioprinting, in realitywe probably won’t see 3D printed, transplantable human organs for many years yet. Internal organs are more complex than simply printing layers of cells into the shape of a kidney or liver. They need to have nerves and blood vessels to function and survive, and bioprinting technology still has a way to go before reaching the required level of complexity.

    The most realistic application of 3D Bioprinting in this decade will most likely be  in drug discovery and development. In the pharma industry, the companies test drugs on animals before the drugs go to expensive clinical trials. However, the human physiology is very different from that of test animals, so what works in an animal, will not necessarily be effective in a person.

    Bioprinting can be used to print a range of 3D culture systems and human tissue models to produce better in vitro testing by generating models with improved physiological relevance and high reproducibility. By applying bioprinting in drug discovery and development organisations can identify ineffective or harmful drugs earlier in the discovery process and shift their resources to more promising drug candidates. They can also reduce the cost of drug development caused by clinical trial failures.

    Organovo’s bioprinted ExVive liver tissues have already proven useful in preclinical toxicology assessment. According to Organovo, the market for liver and kidney in vivo tissue testing is currently valued at close to $3 billion combined1. 3D bioprinting has also been shown to enable the investigation of cancer progression, including tumour heterogeneity, cancer metastasis, and patient specific anticancer drug testing. As bioprinting proves to be a cost-effective and efficient solution, its value in this field is expected to grow from the current $11 million to several hundred millions of dollars over the next decade1.

     

    1SmartTechMarkets (2017) Use Of 3D Bioprinting In Drug Discovery And Cosmetics Testing Expected To Reach $500 Million By 2027 [link]

     

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  4. Biogelx hydrogels utilized to generate a 3D model of a human epiblast

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    An article by MijoSimunovic and colleagues from the Brivanlou and Siggia labs at The Rockefeller University in New York City showcases the use of Biogelx hydrogels in a 3D model of a human epiblast.

    The authors used human embryonic stem cells (hECSs) to create an in vitro 3D model of an epiblast in which size, cell polarity, and gene expression mimicked those of a 10-day human epiblast.

    The epiblast is a polarized epithelium, which will give rise to all cells of the fetus. During gastrulation, the three body axes (anterior-posterior, dorsal-ventral and the left-right axes) are created, establishing the vertebrate body plan. Mouse gastrulation 3D models (called gastruloids) have been previously generated from mouse embryonic stem cells (mESCs), and the morphogenetic movements that establish body symmetry at gastrulation and much of the signaling network underlying these processes have been elucidated in the mouse. However, some of these complex mechanisms are different in humans. Moreover, directly studying human gastrulation and human embryonic development is very challenging due to ethical guidelines (among other considerations) and has been largely unexplored. Therefore, synthetic models of human embryos are needed to address these questions.

    In this work, the authors initallly used Matrigel to culture 3D hESC colonies, however observed the cells differentiated despite the absence of added morphogens. Thus, the researchers concluded that they could not use a Matrigel system to model a pre-gastrulating human epiblast. To improve culture conditions, Simunovic and colleagues utilized Biogelx peptide hydrogel as the 3D matrix.They observed how the 3D hESC colonies displayed “…similar size range, morphology, molecular markers, and tissue polarity that matched our in vitro attached 10-day human embryos”. Furthermore, cells cultured in this 3D model formed a quasi-spherical hollow shell and uniformly expressed key biomarkers like NANOG, SOX2, and OCT4 unlike in pure Matrigel. Additional parameters like the radius of the human epiblast and the surface cell density compared very well with their model epiblast.

    This shows how Biogelx, a peptide hydrogel with defined composition and controlled chemical and mechanical properties proves to be very useful in modelling complex systems which are aim to elucidate intricate processes. Biogelx gels can be utilized either combined with other biomaterials or on their own; especially when using the new functionalized products which are based on the original Biogelx technology, but also contain specific biomimetic sequences relevant to the ECM and basement membrane proteins.

    REFERENCES: Simunovic et al. (2018) Molecular mechanism of symmetry breaking in a 3D model of a human epiblast.bioRxiv 330704.

    You can read the full article here.

     

     

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  5. Disease-driven Changes in the Extracellular Matrix

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    It is widely accepted that cell-based assays traditionally performed in 2D on plastic surfaces are not representative of cells residing in the complex microenvironment of a tissue. This discrepancy is thought to make these systems relatively poor models to predict drug responses, and therefore a contributing factor to the high failure rate in drug discovery. It is believed that 3D cell culture can better reproduce the physiochemical environment of in vivo tissue, providing cells mechanical and biochemical cues like those of native ECM. Therefore, it can provide more physiologically-relevant results, which can ultimately lead to better precision in drug discovery. Despite increased focus, major challenges remain in creating 3D cell-based assays which are reproducible and can recapitulate microenvironmental factors that resemble in vivo tissue as well as disease pathology.

    From the smoothness of brain tissue to the toughness of bone, the mechanical properties of organs vary dramatically. The differences in these environments arise through cells secreting various ECM proteins and macromolecules in different proportionsin addition to intrinsic phenotypic intracellular cell type differences. Whilst such environments are regulated by depositions and degradation of these molecules during homeostasis, some diseases and injuries can disrupt this balancing process. Macromolecules in diseased ECM can be over-represented (upregulated) or reduced (downregulated) compared to levels in healthy tissue, meaning the microenvironment experienced by the cells drastically different in terms of the biochemical composition and in turn, mechanical properties. For example, major components of the microenvironment of the liver are collagen I, II, and IV, laminin, and elastin. During hepatic fibrosis downregulation of elastin is observed alongside an increase in liver stiffness as the disease progresses.

    Similarly, with regards to cancer it is known that most solid tumours are stiffer  than normal tissue, indeed breast cancer is usually screened by detecting hard lumps in the breast. This stiffening is generally accompanied by an increase in deposition of collagen I, II, III, V, and IX during tumour formation. These examples demonstrate that the different properties of the ECM are not independent; rather, they are intertwined. When the ECM stiffens under pathological conditions, its biomechanical properties change, and cells respond by exerting markedly different kinds of force. In addition, matrix stiffening also changes other ECM physical properties and, consequently, directly impacts how migrating cells interact with the ECM.

    Therefore, when developing cell-based models the ability to not only replicate the mechanical properties of the tissue of interest, but also being able to recapitulate the unbalanced ECM composition in diseased/injured tissue is important for providing a more physiologically relevant environment. This will enable amongst other things deeper investigation of the underlying mechanism of disease development, and more predictive pre-clinical assessment on the effects of drugs on both diseased and healthy tissue.

    Employing 3D cell culture matrices which incorporate cell adhesion peptides (CAPs) offers an excellent way to reproduce both healthy tissue- and diseases-specific ECM compositions in a synthetic system. Biogelx offers a range of peptide-based hydrogels which not only allow the stiffness of the final gels to be controlled to match the mechanical properties of specific tissues, but also incorporate biomimetic peptide sequences to mimic key ECM proteins, including fibronectin (RGD), Collagen (GFOGER) and Laminin (IKVAV and YIGSR). Whilst these products are available off the shelf, the simple modular nature of Biogelx technology means that it can be easily customised to your requirements e.g. we can work with you to develop gel formulations incorporating alternative CAPs to remodel a specific tissue microenvironment, or we can optimise the concentration and ratio of CAPs incorporated with your specific application in mind. So, if you are looking to develop more disease-relevant cell culture models, get in touch to discuss our hydrogel customisation service .

     

    References

    1. Forget et. al., Trends Biotechnol., 2018, 36, 4, 372-383
    2. Bataller & D. Brenner, J. Clin. Invest., 2005, 115, 209-218
    3. Lu et. al., J. Cell Biol., 2012, 196, 4, 395–406
    4. Yu et. al.,Trends Cell Biol.,2011, 21, 47–56

     

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