The EGC/PK060399egfr line is a relatively new enteroglial cell (commonly known by the acronym EGC) line initiated by a team of German researchers (A. Ruhl, J. Trotter, and W. Stremmel). The line was established from a myenteric plexus sample from the jejunum of an adult male laboratory rat (Rattus rattus; Sprague-Dawley strain).

Cultured EGC/PK060399egfr cells exhibit adherent growth and strong glial fibrillary acidic protein (GFAP), S-100, and vimentin immunoreactivities. The line does not show signs of Thy-1.1, desmin, smooth muscle alpha-actin, or C3 complement receptor immunoreactivity.

Many details regarding enteroglial cells remain a mystery, but the establishment of the EGC/PK060399egfr line and other EGC lines may eventually help demystify these important components of the enteric nervous system. A. Ruhl and colleagues developed their method of isolating and purifying enteroglial cells from the myenteric plexus specifically with this goal in mind. The technique utilized by the group entailed enzymatic dissociation of myenteric plexus samples, the purification of EGC cells through complement-mediated cytolysis of contaminating cells, transformation via retroviral gene transfer, and characterization of the clones immunohistochemically and by dot-blot analysis.

The cytoskeletal filamentous actin network was labeled in the culture of EGC cells presented in the digital image above with Coumarin conjugated to phalloidin (yielding blue emission). Mitochondria and nuclei were also visualized in the enteroglial cells with MitoTracker Red CMXRos and SYTOX Green, respectively. 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.

Triple Fluorophore Labeling of Enteroglial Cells with MitoTracker Red CMXRos, BODIPY FL, and Hoechst 33258 - The culture of EGC cells presented in the digital image in this section was triple-labeled for mitochondria, filamentous actin, and nuclear DNA with MitoTracker Red CMXRos, BODIPY FL conjugated to phalloidin, and Hoechst 33258, respectively.

Localizing the Golgi Complex in EGC Cell Cultures with Wheat Germ Agglutinin - A culture of rat jejunum enteroglial cells was labeled with wheat germ agglutinin (WGA) conjugated to Oregon Green 488. WGA, which selectively binds to N-acetylglucosamine and N-acetylneuraminic (sialic acid) residues, is well suited for staining the Golgi network in fixed cells since a number of proteins and lipids found in the Golgi apparatus are glycosylated.

F-Actin and Mitochondria Distribution in Rat Jejunum Enteroglial Cells - MitoTracker Red CMXRos and Alexa Fluor 488 conjugated to phalloidin were utilized to label the featured culture of EGC cells, targeting the mitochondrial network and filamentous actin, respectively. The culture was counterstained for DNA in the cell nucleus with Hoechst DAPI.

EGC Cell Culture Labeled with Soybean Agglutinin - The adherent culture of enteroglial cells illustrated in this section was labeled with Alexa Fluor 488 conjugated to soybean agglutinin, a lectin isolated from Glycine max that has saccharide binding sites that recognize and bind terminal alpha- and beta-N-acetylgalactosamine and galactopyranosyl oligosaccharides. In addition, the cells were labeled with Alexa Fluor 568 conjugated to phalloidin and DAPI, targeting the cytoskeletal F-actin network and DNA, respectively.

Visualizing Mitochondria, Filamentous Actin, and DNA in Rat Enteroglial Cell Cultures - The popular triple fluorophore combination of MitoTracker Red CMXRos, BODIPY FL conjugated to phalloidin, and Hoechst 33258 was used to label the adherent log phase culture of rat jejunum enteroglial cells presented in this section for mitochondria, the filamentous actin network, and nuclear DNA.

Cultured EGC Cells with MitoTracker Red CMXRos, Alexa Fluor 488, and DAPI - The mitochondrial and cytoskeletal F-actin networks were visualized in a culture of rat jejunum myenteric plexus enteroglial cells by labeling them with MitoTracker Red CMXRos and Alexa Fluor 488 conjugated to phalloidin, respectively. Cell nuclei were counterstained with DAPI.