The Madin-Darby ovine kidney (MDOK) cell line was initiated in the 1950s from kidney tissue excised from a normal adult male sheep (Ovis aries). The cells exhibit epithelial characteristics and are useful in virology and veterinary virology research.

MDOK cells are known to be susceptible to a variety of viruses, including sheep bluetongue virus, vesicular stomatitis (Indiana and New Jersey strains), and infectious bovine rhinotracheitis. Similar to other epithelial lines, MDOK cells experience significant contact inhibition of migration in culture. The cells grow adherently to both glass and polymer surfaces.

The behavior of Madin-Darby ovine kidney cells in culture is thought to reflect the behavior of epithelial cells in the body. In vivo, epithelial cells are generally found bound to one another in sheets that line the internal and external surfaces of the body and its organs. A variety of specialized junctions form between the cells, including a type of junction unique to epithelia called a tight junction. The integrity of epithelial sheets is extremely important because they primarily serve as barriers against diffusion. Some epithelial cells also function in absorptive or secretory roles as well.

Three fluorescent probes were utilized to label the Madin-Darby ovine kidney cell culture depicted in the digital image above, including MitoTracker Red CMXRos (mitochondria), Alexa Fluor 633 conjugated to phalloidin (F-actin), and SYTOX Green (nuclei). Images were recorded in grayscale with a 12-bit digital camera coupled to either a Nikon E-600 or Eclipse 80i microscope equipped with bandpass emission fluorescence filter optical blocks. During the processing stage, individual image channels were pseudocolored with RGB values corresponding to each of the fluorophore emission spectral profiles.

Distribution of Nonmuscle Myosin II and F-Actin in MDOK Cell Cultures - A fixed and permeabilized monolayer culture of MDOK cells was treated with mouse anti-nonmuscle myosin II (heavy chain) monoclonal primary antibodies followed by goat anti-mouse secondary antibodies (IgG) conjugated to Texas Red-X. Oregon Green 488 conjugated to phalloidin was included in the secondary cocktail to target the filamentous actin network.

Localizing the Golgi Complex and the Nucleus in Ovine Kidney Cells - The proximity of nuclei and the Golgi complex in the MDOK cell culture illustrated in this section was probed in a double immunofluorescence experiment with mouse anti-histones (pan) and rabbit anti-giantin primary antibodies. The antibody targets were visualized with goat secondary antibodies conjugated to Texas Red and Alexa Fluor 488, respectively. In addition, the F-actin cytoskeletal framework was labeled with Alexa Fluor 350 conjugated to phalloidin.

Triple Fluorophore Labeling of MDOK Cells with Alexa Fluor 633, MitoTracker Red CMXRos, and SYTOX Green - The filamentous actin components of the cytoskeletal framework were labeled in a culture of MDOK epithelial cells with Alexa Fluor 633 conjugated to phalloidin. Mitochondria and DNA in the cell nucleus were also probed with MitoTracker Red CMXRos and SYTOX Green, respectively.

Proximity of the Nucleus and Mitochondria in Madin-Darby Ovine Kidney Cell Cultures - The mitochondrial network was targeted in a culture of Madin-Darby ovine kidney cells with MitoTracker Red CMXRos. In addition, cell nuclei were counterstained with Hoechst 33258.

Targeting Histones and Giantin in Sheep Kidney Cells with Immunofluorescence - Nuclear histone proteins were targeted in the culture of MDOK cells presented in this section with mouse anti-histone (pan) monoclonal antibodies, which were imaged with goat anti-mouse Fab fragments conjugated to Texas Red (labeling the nucleus). The specimen was simultaneously labeled for the Golgi complex with Alexa Fluor 488 conjugated to goat secondary antibodies that target rabbit anti-giantin. Alexa Fluor 350 conjugated to phalloidin was utilized to counterstain the cytoskeletal F-actin network.