In vitro 3D cancer models that provide a more accurate representation of disease in vivo are urgently needed to improve our understanding of cancer pathology and to develop better cancer therapies. However, development of 3D models that are based on manual ejection of cells from micropipettes suffer from inherent limitations such as poor control over cell density, limited repeatability, low throughput, and, in the case of coculture models, lack of reproducible control over spatial distance between cell types (e.g., cancer and stromal cells). In this study, we build on a recently introduced 3D model in which human ovarian cancer (OVCAR-5) cells overlaid on Matrigel™ spontaneously form multicellular acini. We introduce a high-throughput automated cell printing system to bioprint a 3D coculture model using cancer cells and normal fi broblasts micropatterned on Matrigel™. Two cell types were patterned within a spatially controlled microenvironment (e.g., cell density, cell-cell distance) in a high-throughput and reproducible manner; both cell types remained viable during printing and continued to proliferate following patterning. This approach enables the miniaturization of an established macro-scale 3D culture model and would allow systematic investigation into the multiple unknown regulatory feedback mechanisms between tumor and stromal cells and provide a tool for high-throughput drug screening. Copyright © 2011 WILEY-VCH Verlag GmbH & Co.

KGaA, Weinheim. Schematic of a high-throughput ejector platform composed of a computerized stage and two ejectors. Dual ejector heads are used to eject different cell types simultaneously, i.e., cancer cells (OVCAR-5) and fibroblasts (MRC-5). (a) The platform is installed in a sterile hood to prevent contamination using HEPA filters. (b) The platform consists of an automated xyz stage and nanoliter dispensing valves controlled by a pulse generator.

Overcome chemoresistance in heterocellular 3D tumor models (Conference Presentation) A major. Imran Rizvi at University of North Carolina at Chapel Hill.

The position of substrate and droplet generator are synchronized and programmed through predefined control commands. Characterization of the high-throughput cell patterning platform (60 μs valve opening duration and 34.5 kPa nitrogen gas pressure). (a) Droplet positioning accuracy. The positioning error of two droplets is measured by the difference between programmed distance (D program) and actual printed distance of droplets after patterning (D actual); R 2=0.9932. (b) Droplet size distribution, measured in liquid nitrogen, was 510 ± 26 μm. Droplet size was obtained from 51 droplets.

Asi front desk download crack windows 10. (c) Number of cells per droplet for OVCAR-5 and MRC-5. (d) Percent viability of OVCAR-5 and MRC-5 coculture after printing (4 h) and at day 3 (72 h) with respect to flask cell viability ( n=4). F) Live/dead staining to calculate percent viability at (d). Scale bars, 250 μm.

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