The myoblast cell line A-10 was established from tissue excised from the medial layer of the thoracic aorta of a rat embryo (Rattus norvegicus; DB1X strain). A-10 cells are frequently utilized in medical research, especially in studies of hypertension and other conditions and diseases potentially associated with smooth muscle cells.

A-10 cells grow adherently to glass and plastic surfaces in culture. When cultures of the cells reach the stationary phase of the growth cycle, they produce spontaneous action potentials and experience increased enzymatic activity of myokinase and creatine phosphokinase. The cellular products of A-10 cells also include myosin, the protein that provides the elastic and contractile properties of muscle.

Many researchers are employing A-10 cells in hypertension studies in an attempt to better understand the condition. One potential cause of hypertension, commonly referred to as high-blood pressure, is increased smooth muscle cell size and local proliferation. More specifically, additional smooth muscle in the walls of the arteries increases the thickness of the walls of the blood vessels and may result in their increased constrictive capacity. These developments can lead to amplified lumen constriction and increased resistance, effectively reducing blood flow. The cardiovascular system is thought to naturally respond to decreased blood flow by elevating blood pressure so that the supply of blood to body tissues does not diminish.

The A-10 myoblast cells presented in the digital image above were resident in an adherent culture stained for F-actin with Alexa Fluor 488 conjugated to phalloidin (green fluorescence), and for nuclear DNA with the bis-benzimidazole dye Hoechst 33258 (blue fluorescence). In addition, the culture was immunofluorescently labeled with Alexa Fluor 568 conjugated to goat secondary antibodies that target mouse anti-PDI (protein disulfide isomerase) primary antibodies (red fluorescence). 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.

Labeling Rat Thoracic Aorta Cells for PDI and PMP 70 with Double Immunofluorescence - In a double immunofluorescence experiment, the culture of rat thoracic aorta cells illustrated in this section was fixed, permeabilized, blocked with 10 percent normal goat serum, and treated with a cocktail of mouse anti-PDI (protein disulfide isomerase) and rabbit anti-PMP 70 (peroxisomal membrane protein) primary antibodies, followed by goat anti-mouse and anti-rabbit secondary antibodies (IgG) conjugated to Alexa Fluor 568 and Alexa Fluor 750, respectively.

Distribution of Histones and Peroxisomal Membrane Proteins in Rat Thoracic Aorta Myoblasts - The adherent monolayer culture of rat thoracic aorta cells presented in this section was immunofluorescently labeled with mouse anti-histones (pan) and rabbit anti-PMP 70 (peroxisomal membrane protein) primary antibodies. Treatment with goat anti-mouse and anti-rabbit secondary antibodies (IgG) conjugated to Texas Red and Alexa Fluor 488 was utilized to visualize the proteins.

Visualizing Vinculin and Giantin in A-10 Cell Cultures - Focal adhesions and the Golgi complex were visualized in an adherent monolayer culture of A-10 cells by immunofluorescent treatment with mouse anti-vinculin primary antibodies and rabbit primary antibodies to giantin (a protein resident in the Golgi complex of mammalian cells) followed by goat anti-mouse Fab fragments conjugated to Alexa Fluor 750 and goat anti-rabbit secondary antibody fragments (heavy and light chain) conjugated to Alexa Fluor 568.

Rat Thoracic Aorta Cytoskeletal Intermediate Filament and Actin Networks - The proximity of intermediate filaments and the cytoskeletal filamentous actin network was visualized by treating the fixed and permeabilized culture of rat thoracic aorta cells presented in this section with mouse anti-vimentin primary antibodies followed by goat anti-mouse secondary antibodies (IgG) conjugated to Texas Red-X. F-actin was subsequently labeled with Alexa Fluor 350 conjugated to phalloidin, and the nuclei were counterstained with SYTOX Green.

Focal Adhesion Sites in A-10 Myoblast Cells - A culture of A-10 cells was immunofluorescently labeled with primary anti-vinculin mouse monoclonal antibodies followed by goat anti-mouse Fab fragments conjugated to Cy3. In addition, the specimen was simultaneously stained for DNA with the ultraviolet-absorbing probe DAPI, and for the cytoskeletal filamentous actin network with Alexa Fluor 488 conjugated to phalloidin.

Five Color Fluorescence in A-10 Myoblast Cells - A culture of A-10 cells was immunofluorescently labeled with a combination of primary anti-vinculin mouse and anti-giantin rabbit monoclonal antibodies followed by goat anti-mouse Fab fragments conjugated to Alexa Fluor 750 and goat anti-rabbit secondary antibodies conjugated to Alexa Fluor 568. In addition, the specimen was simultaneously stained for DNA with the ultraviolet-absorbing probe Hoechst 33342, for mitochondria with MitoTracker Deep Red 633, and for the cytoskeletal filamentous actin network with Alexa Fluor 488 conjugated to phalloidin.