In vitro models that capture the complexity of in vivo tissue and organ behaviors in a scalable and easy-to-use format are desirable for drug discovery. To address this, we have developed a bioreactor that fosters maintenance of 3D tissue cultures under constant perfusion and we have integrated multiple bioreactors into an array in a multiwell plate format. All bioreactors are fluidically isolated from each other. Each bioreactor in the array contains a scaffold that supports formation of hundreds of 3D microscale tissue units. The tissue units are perfused with cell culture medium circulated within the bioreactor by integrated pneumatic diaphragm micropumps. Electronic controls for the pumps are kept outside the incubator and connected to the perfused multiwell by pneumatic lines. The docking design and open-well bioreactor layout make handling perfused multiwell plates similar to using standard multiwell tissue culture plates. A model of oxygen consumption and transport in the circulating culture medium was used to predict appropriate operating parameters for primary liver cultures. Oxygen concentrations at key locations in the system were then measured as a function of flow rate and time after initiation of culture to determine oxygen consumption rates. After seven days in culture, tissue formed from cells seeded in the perfused multiwell reactor remained functionally viable as assessed by immunostaining for hepatocyte and liver sinusoidal endothelial cell (LSEC) phenotypic markers.

Assessment of drug–liver interactions is an integral part of predicting the safety profile of new drugs. Existing model systems range from in vitro cell culture models to FDA-mandated animal tests. Data from these models often fail, however, to predict human liver toxicity, resulting in costly failures of clinical trials. In vitro screens based on cultured hepatocytes are now commonly used in early stages of development, but many toxic responses in vivo seem to be mediated by a complex interplay among several different cell types. We discuss some of the evolving trends in liver cell culture systems applied to drug safety assessment and describe an experimental model that captures complex liver physiology through incorporation of heterotypic cell–cell interactions, 3D architecture and perfused flow. We demonstrate how heterotypic interactions in this system can be manipulated to recreate an inflammatory environment and apply the model to test compounds that potentially exhibit idiosyncratic drug toxicity. Finally, we provide a perspective on how the range of existing and emerging in vitro liver culture approaches, from simple to complex, might serve needs across the range of stages in drug discovery and development, including applications in molecular therapeutics.

This paper characterizes a bi-directional pneumatic diaphragm micropump and presents a model for performance of an integrated fluidic capacitor. The fluidic capacitor is used to convert pulsatile flow into a nearly continuous flow stream. The pump was fabricated in acrylic using a CNC mill. The stroke volume of the pump is ∼1 µL. The pump is self-priming, bubble tolerant and insensitive to changes in head pressure and pneumatic pressure within its operating range. The pump achieves a maximum flow rate of 5 mL min−1 against zero head pressure. With pneumatic pressure set to 40 kPa, the pump can provide flow at 2.6 mL min−1 against a head pressure of 25 kPa. A nonlinear model for the capacitor was developed and compared with experimental results. The ratio of the time constant of the capacitor to the cycle time of the pump is shown to be an accurate indicator of capacitor performance and a useful design tool.

The vast majority of cancer mortalities result from distant metastases. The metastatic microenvironment provides unique protection to ectopic tumors as the primary tumors often respond to specific agents. Although significant interventional progress has been made on primary tumors, the lack of relevant accessible model in vitro systems in which to study metastases has plagued metastatic therapeutic development - particularly among micrometastases. A real-time, all-human model of metastatic seeding and cancer cells that recapitulate metastatic growth and can be probed in real time by a variety of measures and challenges would provide a critical window into the pathophysiology of metastasis and pharmacology of metastatic tumor resistance. To achieve this we are advancing our microscale bioreactor that incorporates human hepatocytes, human nonparenchymal liver cells, and human breast cancer cells to mimic the hepatic niche in three dimensions with functional tissue. This bioreactor is instrumented with oxygen sensors, micropumps capable of generating diurnally varying profiles of nutrients and hormones, while enabling real-time sampling. Since the liver is a major metastatic site for a wide variety of carcinomas and other tumors, this bioreactor uniquely allows us to more accurately recreate the human metastatic microenvironment and probe the paracrine effects between the liver parenchyma and metastatic cells. Further, as the liver is the principal site of xenobiotic metabolism, this reactor will help us investigate the chemotherapeutic response within a metabolically challenged liver microenvironment. This model is anticipated to yield markers of metastatic behavior and pharmacologic metabolism that will enable better clinical monitoring, and will guide the design of clinical studies to understand drug efficacy and safety in cancer therapeutics. This highly instrumented bioreactor format, hosting a growing tumor within a microenvironment and monitoring its responses, is readily transferable to other organs, giving this work impact beyond the liver.

The most complex types of nitrogen compound in beer (here termed "proteoses") are, when not combined with polyphenols, highly surface active and hence concentrate in the foam.With unhopped beer, the foam is of the "liquidviscous" type, which eventually drains and collapses leaving no residual solids. The iso-a-acids in hopped beers are also surface-active, so they too are con centrated In the foam.At the corresponding concentration in bulk solution the solubility limits of some isohumulates* are exceeded.Thus we believe that solids are similarly formed in the bubble films, so that these are reinforced and stiffened to give "cling".One consequence of the increased concentration of iso-a-acids in the foam is that there can be precipitation of iron, nickel, cobalt and zinc isohumulates. This results in concentration of these metals in foam.The most significant finding Is that the concentration of both proteoses and iso-a-acids in the foam exceeds the bulk solubility limits and results in the formation of proteose-isohumulate salts which make the main contribution to the solid reinforcement of bubble films.These findings make it possible to explain satisfactorily "lacing" or "cling" and also isohumulone losses (a) during boiling and trub separation, (b) during post-fermentation bittering, and (c) during any subsequent foam for mation. An explanation is also provided why losses of bitterness and of head retention are linked.Consequently, linked recovery of both bitterness and head retention can occur, as has been found is some recent fermentation im provements, such as continuous fermentation.