Many factors contribute to the creation and maintenance of a realistic environment for cell growth in vitro, e.g. the consistency of the growth medium, the addition of supplements, and the surface on which the cells grow. The nature of the surface on which cells are cultured plays an important role in their ability to attach, proliferate, migrate and function. Components of the extracellular matrix (ECM) are often used to coat glass or plastic surfaces to enhance cell attachment in vitro. Fragments of ECM molecules can be immobilised on surfaces in order to mimic the effects seen by whole molecules. In this study we evaluate the application of a novel technology for the immobilisation of functional domains of known ECM proteins in a controlled manner on a surface. By examining the adherence of cultured PC12 cells to alternative growth surfaces, we show that surfaces coated with motifs from collagen I, collagen IV, fibronectin and laminin can mimic surfaces coated with the corresponding whole molecules. Furthermore, we show that the adherence of cells can be controlled by modifying the hydropathic properties of the surface to either enhance or inhibit cell attachment. Collectively, these data demonstrate the application of a new technology to enable optimisation of cell growth in the tissue culture laboratory.
The development is reported of an ultra-rapid, point-of-care diagnostic device which harnesses surface acoustic wave (SAW) biochips, to detect HIV in a finger prick of blood within 10 seconds (samplein-result-out). The disposable quartz biochip, based on microelectronic components found in every consumer smartphone, is extremely fast because no complex labelling, amplification or wash steps are needed. A pocket-sized control box reads out the SAW signal and displays results electronically. High analytical sensitivity and specificity are found with model and real patient blood samples. The findings presented here open up the potential of consumer electronics to cut lengthy test waiting times, giving patients on the spot access to potentially life-saving treatment and supporting more timely public health interventions to prevent disease transmission.Ebola and Zika viruses offer a stark reminder that infectious diseases rank among the gravest threats to human health, and can spread rapidly and unpredictably. New infections will continue to emerge each year, and old enemies re-emerge, increasingly with acquired-drug resistance (e.g. gonorrhoea and HIV). Rapid diagnosis plays a crucial role in any outbreak situation, empowering patients to gain faster access to potentially life-saving treatment, and informing prevention strategies to protect the wider public. However, routine diagnostic tests based on enzyme linked immunosorbent assays (ELISAs) and polymerase chain reaction (PCR) are confined to centralized laboratories often requiring large, sophisticated, costly instrumentation and highly trained staff. Inherent delays occur between taking samples, conveying them to the laboratory, waiting for results to come back and subsequent follow up appointments 1-3 . This means that a patient often has to make multiple visits to a clinic in order to receive treatment, potentially over long distances. This delays prescribing of treatment with increased risk of suffering, mortality, and also incorrect prescription of antimicrobials.Recent policy drivers aim to widen access to testing using so called 'rapid' point-of-care tests (POCT) but the performance and implementation of these tests still remain a challenge 4 . The most common tests based on lateral flow technology are still relatively slow, requiring a 10-20 minute waiting time for results 5 . This exceeds a typical doctor's appointment (8-10 mins in the UK 6 ) necessitating changes to patient pathways within a clinic with additional on-costs and staffing implications. It is also notoriously difficult to interpret a faint lateral flow test line by eye, particularly for non-experts (e.g. self-testers) 1 . Those tests that are currently available are insensitive to recent (acute) infections 7 and lack the ability to automatically capture test results electronically, risking an incorrect reading, missed opportunities to link patients to care pathways and potential data loss for public health (e.g. during an Ebola outbreak) 8 . Alternatively, uneccesary treatment may be initi...
The interaction between cells and the extracellular matrix (ECM) is essential during development. To elucidate the function of ECM proteins on cell differentiation, we developed biomimetic surfaces that display specific ECM peptide motifs in a controlled manner. Presentation of ECM domains for collagen, fibronectin, and laminin influenced the formation of neurites by differentiating PC12 cells. The effect of these peptide sequences was also tested on the development of adult neural stem/progenitor cells. In this system, collagen I and fibronectin induced the formation of beta-III-tubulin positive cells, whereas collagen IV reduced such differentiation. Biomimetic surfaces composed of multiple peptide types enabled the combinatorial effects of various ECM motifs to be studied. Surfaces displaying combined motifs were often predictable as a result of the synergistic effects of ECM peptides studied in isolation. For example, the additive effects of fibronectin and laminin resulted in greater expression of beta-III-tubulin positive cells, whereas the negative effect of the collagen IV domain was canceled out by coexpression of collagen I. However, simultaneous expression of certain ECM domains was less predictable. These data highlight the complexity of the cellular response to combined ECM signals and the need to study the function of ECM domains individually and in combination.
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