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drugs and Cell culture

Peterer
29.05.2018

Content:

  • drugs and Cell culture
  • Comparison of 2D- and 3D-culture models as drug-testing platforms in breast cancer
  • Materials and Methods
  • Cell culture is among the most widely used laboratory approaches probably because of the diverse aspects it covers and the relatively short period of time it. In this review, we will provide an overview on the most common 3D cell culture techniques, address the opportunities they provide for both drug. Toxicological screening of drug candidates early in development with in vitro cell culture systems is therefore of relevance. In contrast to animal studies, in vitro.

    drugs and Cell culture

    Author information Article notes Copyright and License information Disclaimer. Cell culture, Drug development. This article has been cited by other articles in PMC. Footnotes Peer review under responsibility of King Saud University. Open in a separate window. Chapter 9 — best practice in toxicological pathology. A spotlight on chemical constituents and pharmacological activities of Nigella glandulifera Freyn et Sint seeds. Is mapping borders between pharmacology and toxicology a necessity?

    Psychiatric neural networks and neuropharmacology: Targeting G protein coupled receptor-related pathways as emerging molecular therapies. Shorter and sturdier bridges between traditional Chinese medicines and modern pharmacology. Chapter 3 — natural product chemistry and cancer drug discovery. Cancer Drug Design and Discovery. Academic Press; San Diego: Chapter 6 — toxicogenomics — a drug development perspective.

    Genomic Biomarkers for Pharmaceutical Development. The fall and rise of pharmacology — Re- defining the discipline? Articles from Saudi Pharmaceutical Journal: SPJ are provided here courtesy of Elsevier.

    Support Center Support Center. Please review our privacy policy. The past decades have witnessed significant efforts toward the development of three-dimensional 3D cell cultures. Today, 3D cell cultures are emerging not only as a new tool in early drug discovery, but also as potential therapeutics to treat disease.

    Cell culture is a fundamental technique in both medical research and drug discovery, and for decades, two-dimensional 2D culture has been the preferred method, due to the ease with which cell monolayers can be induced to proliferate on planar surfaces. However, early pioneering work in cell culture utilised tissue explants grown in three-dimensions 3D.

    It was evident that such 3D cultures maintained many attributes that resembled cell growth and differentiation in vivo 1.

    Today, the limitations of 2D culture, the development of advanced laboratory products and sophisticated detection instrumentation and analytic software, as well as the emergence of stems cells as powerful research tools, has led to the growing adoption of 3D culture models in many phases of drug discovery, including target validation, lead identification and preclinical optimisation.

    In the past, 3D cell culture models were mostly developed for oncology research, given that tumours exist as 3D entities in vivo and, therefore, should be better approximations of the tumour microenvironment 2.

    Consequently, several 3D cell tumour models now exist, from multicellular layers on porous membranes coated with collagen, to matrix-embedded cultures, hollow fibre bioreactors and multicellular spheroids.

    A more recent approach, yet to have a major impact on drug discovery, has been the growing use of dish-based organogenesis, using technologies from stem cell research and mixed cell culture techniques 3. This approach has a high degree of physiological relevance and results in the development of organoids with realistic micro-anatomy. Taken together, 3D cell culture techniques are no longer confined to the research space, but are emerging as a powerful new tool in preclinical drug discovery.

    Recent advances in cell biology, microfabrication techniques and tissue engineering have enabled the development of a wide range of 3D cell culture technologies.

    These include multicellular spheroids, organoids, scaffolds, hydrogels, organson-chips, and 3D bioprinting, each with its own advantages and disadvantages, see Table 1 for a summary.

    Furthermore, since patient-specific cells can also be grown in 3D, the exciting possibility exists for drug discovery to be undertaken on cells with very precise pathophysiologies. Finally, 3D cell culture is growing in the area of bioproduction, notably as a means to scale-up abundant and reproducible numbers of cells as potential therapeutics.

    Cellular assays are routinely used in compound screening and optimisation, with growing evidence showing that compound library hits and optimised leads translate into better candidates for clinical evaluation 4,5. The majority of cell-based screening is performed using 2D culture technologies, due in part to the demands of the automation and detection instrumentation in use.

    However, cells grown in 3D better reflect drug-target interactions in vivo 6,7. Additionally, drug sensitivity in 3D culture models differs markedly from that obtained using 2D culture Figure 1.

    In particular, cell-based screening technology has been pivotal in the area of imaging technologies as used in high content screening HCS assays. Further, many of the current technologies that enable 3D cell culture also support co-culture conditions, allowing for multiple cell types to be integrated into a 3D model that more closely mimics the in vivo microenvironment.

    These co-culture models are important for drug discovery, as the presence of certain cell types within tumour spheroids can greatly shift drug responsiveness Figure 2. To date, more than types of matrices and scaffolds have been developed, most of which are optimised to the growth of the specific cells under investigation.

    Naturally-derived ECMs are widely used in 3D cell culture. These basement membrane hydrogels can provide the appropriate microenvironment needed for morphogenesis and organogenesis of cells possessing intrinsic developmental programmes. Immortalised cell lines and tissue fragments form structures that recapitulate key tissue features when embedded in ECM gels and exposed to appropriate growth factors. Stem cell-derived organoids have been developed from embryonic stem cells ESCs , induced pluripotent stem cells IPSCs or from primary stem cells purified from organs3.

    The possibility now exists for patient-derived organoids that potentially enable personalised approaches to identify the mechanisms underlying human diseases, and to evaluate the efficacy and predict toxic potential of drugs prior to administration. This transformative approach may help identify the best therapies for individual cancer patients over the course of their disease.

    Cellular spheroids embedded in ECMs present several features, including a defined geometry, optimal physiological cell-cell and cell-ECM interactions, and better gradients of nutrients, growth factors and oxygen, upon which transport occurs for several hours or even days 7. These attributes facilitate screening assays for compounds to modulate tumour growth, invasion and angiogenesis. Cellular spheroids can be generated from many types of cells; those formed include embryoid bodies, mammospheres, tumour spheroids, hepatospheres and neurospheres.

    Naturally-derived hydrogels for 3D culture comprise proteins and other ECM components, including collagen, laminin and fibrin.

    A major advantage of 3D culture using a naturally-derived hydrogel is that the protocols are robust and simple. However, they have certain disadvantages due to their origin as undefined, complex material of variable compositions. Furthermore, naturally-derived hydrogels may lack the mechanical properties provided by endogenous ECMs. Finally, their nonhuman origin can preclude their use in human regenerative or transplantation therapies.

    The microenvironment is clearly critical to complete organ development; but this feature has, nonetheless, been difficult to reconstitute completely in many 3D cultures.

    The development of synthetic structures or scaffolds using naturally-derived ECM, synthetic hydrogels or other biocompatible materials may address this issue, as they are designed to either replace or complement naturallyderived ECMs with clinical-grade materials. Tissue organoids in 3D ECMs have been developed for mammary, stomach, intestinal, liver, brain, salivary, kidney, lung and pancreatic ductal epithelium 3, However, the thick ECM gel can limit optical imaging and cell recovery is more complex.

    To overcome some of these limitations, several lab consumables and methods have been developed to take advantage of spontaneous cell-cell interactions, which occur when cells are in an environment that promotes greater attraction towards each other than to any available surface. Of the techniques developed to take advantage of the phenomenon of cell aggregation, the hanging drop 13 and low-attachment methods 14 Figure 3 are widely used due to their compatibility with automated screening instrumentation and detection systems.

    Cell therapy and tissue engineering not only offer new hope for patients with injuries, end-stage organ failure, or other clinical issues, but will eventually transform our lives. However, it is becoming clear that realising the full potential of cell therapy and tissue engineering requires advances in cell culture technologies to meet the demand in quantity, quality and process robustness for commercialisation and clinical trials.

    Stem cells are widely used as a cell source for regenerative medicine and cell therapy applications. However, conventional 2D culture techniques, in combination with the current best practice, may be ineffective in the expansion of stem cells for clinical applications. This is reflected by the fact that 2D cultures are inadequate to reproduce the in vivo microenvironment of stem cells In addition, clinical observations show that the beneficial effects of stem cell-based therapeutics seen in initial small-scale clinical studies are often not validated by large, randomised clinical trials 17, In fact, mesenchymal stem cells MSCs often decrease their replicative ability, colony forming efficiency and differentiation capabilities over time when culturing and passaging in 2D adherent monolayer 19, In contrast, MSCs cultured in spheroids display a morphology that is significantly different from 2D culture Furthermore, compared with 2D culture, MSCs cultured in spheroids have different gene expression patterns, with up-regulation of many genes that are associated with hypoxia, angiogenesis, inflammation, stress response and redox signalling Spheroid cultures have been reported to improve the efficacy of MSC-based therapeutics.

    Compared with 2D cultures, MSC spheroid cultures were also found to have additional benefits, such as enhanced anti-inflammatory and tissue regenerative and reparative effects, as well as better post-transplant survival of MSCs Furthermore, compared with 2D cultured cells, spheroids of human adiposederived MSCs produced higher levels of ECM proteins, exhibited stronger antiapoptotic and antioxidative capacities and increased the paracrine secretion of cytokines.

    When considering transplantation, organoids could provide a source of autologous tissue, as organoid research advances rapidly. For instance, renal organoids derived from pluripotent stem cells were successfully transplanted under the renal capsules of adult mice Here, the organoid reconstituted the 3D structures of the kidney in vivo, including glomeruli with podocytes and renal tubules with proximal and distal regions and clear lumina.

    Comparison of 2D- and 3D-culture models as drug-testing platforms in breast cancer

    Although, many types of in vitro assays are conducted during drug development, use of cell cultures is the most reliable one. Two-dimensional (2D) cell cultures. Today, 3D cell cultures are emerging not only as a new tool in early drug discovery, but also as potential therapeutics to treat disease. Cell culture is a. PDF | Cell culture systems are one of the widely used systems implicated in drug testing and drug development. Recombinant cell lines.

    Materials and Methods



    Comments

    Zologis

    Although, many types of in vitro assays are conducted during drug development, use of cell cultures is the most reliable one. Two-dimensional (2D) cell cultures.

    zadanters

    Today, 3D cell cultures are emerging not only as a new tool in early drug discovery, but also as potential therapeutics to treat disease. Cell culture is a.

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