Drug discovery programmes require accurate in vitro systems for drug screening and testing. Traditional cell culture makes use of 2D (two-dimensional) surfaces for ex vivo cell growth. In such environments, cells are forced to adopt unnatural characteristics, including aberrant flattened morphologies. Therefore there is a strong demand for new cell culture platforms which allow cells to grow an...

There are two main methods for preparing the PuraMatrix (PM) as a three-dimensional (3-D) scaffold for supporting the cells including surface plating and encapsulation. In the surface-plating procedure, the scaffold is created by adding an ionic agent such as medium to PM. After adjusting the PH by changing the medium, cells are seeded on top of the scaffold. In the encapsulation method, PM and...

In renal collecting duct (CD) principal cells (PCs), vasopressin (VP) acts through its receptor, V2R, to increase intracellular cAMP leading to phosphorylation and apical membrane accumulation of the water channel aquaporin 2 (AQP2). The trafficking and function of basolaterally located AQP2 is, however, poorly understood. Here we report the successful application of a 3-dimensional Madin-Darby...

Thixotropic materials, which become less viscous under stress and return to their original state when stress is removed, have been used to deliver gel-cell constructs and therapeutic agents. Here we show that a polymer-silica nanocomposite thixotropic gel can be used as a three-dimensional cell culture material. The gel liquefies when vortexed--allowing cells and biological components to be add...

Characterizing how cells in three-dimensional (3D) environments or natural tissues respond to biophysical stimuli is a longstanding challenge in biology and tissue engineering. We demonstrate a strategy to monitor morphological and mechanical responses of contractile fibroblasts in a 3D environment. Cells responded to stretch through specific, cell-wide mechanisms involving staged retraction an...

BioSig3D is a computational platform for high-content screening of three-dimensional (3D) cell culture models that are imaged in full 3D volume. It provides an end-to-end solution for designing high content screening assays, based on colony organization that is derived from segmentation of nuclei in each colony. BioSig3D also enables visualization of raw and processed 3D volumetric data for qua...

Methods. Cells and reagents We purchased MDA-MB-231 cells from ATCC and primary IMR90, HDF and primary HUVEC cells from Lonza. MDA-MB-231, IMR-90, HDF and LLC cells were cultured in Eagle’s minimal Essential medium (MEM) from Lonza supplemented with 10% fetal bovine serum (FBS) and 1% GlutaMax (both from Invitrogen). Primary HUVEC cells were cultured in endothelial growth medium (EGM-2, Lonza)....

Background: MCF-10A cells are near diploid and normal human mammary epithelial cells. In three-dimensional reconstituted basement membrane culture, they undergo a well-defined program of proliferation, differentiation, and growth arrest, forming acinar structures that recapitulate many aspects of mammary architecture in vivo. The pre-malignant MCF-10AT cells and malignant MCF-10CA1a lines were ...

Hepatocytes self-assemble in culture to form compacted spherical aggregates, or spheroids, that mimic the structure of the liver by forming tight junctions and bile canalicular channels. Hepatocyte spheroids thus resemble the liver to a great extent. However, liver tissue contains other cell types and has bile ducts and sinusoids formed by endothelial cells. Reproducing 3-D co-culture in vitro ...

Cells, grown as monolayers (2D models), are routinely used as initial model systems for evaluating the effectiveness and safety of libraries of molecules with potential as therapeutic drugs. While this initial screening precedes preclinical animal studies before advancing to human clinical trials, cultured cells frequently determine the initial, yet crucial, 'stop/go' decisions on the progressi...