Isolation of enteric glia and establishment of transformed enteroglial cell lines from the myenteric plexus of adult rat.
Although enteroglial cells (EGCs) may play a key role in the inflammatory response of the enteric nervous system, little is known about their immunophysiological properties. To facilitate further characterization of enteric glia, we have developed a novel method to isolate and purify EGCs from the myenteric plexus. Myenteric plexus preparations were enzymatically dissociated and EGCs purified by complement-mediated cytolysis of contaminating cells and transformed by retroviral gene transfer. Primary and transformed cells were characterized immunohistochemically and by dot-blot analysis. Functionally, c-fos mRNA expression was assessed in primary and transformed enteroglial cells. All cells displayed robust glial fibrillary acidic protein, S-100 and vimentin immunoreactivities, but no Thy-1.1, desmin, smooth muscle alpha-actin or C3 complement receptor immunoreactivity. This confirmed their enteroglial lineage and excluded contamination with other cell types. Both primary and transformed EGCs displayed little constitutive c-fos mRNA expression. This, however, could be upregulated by various stimuli, including proinflammatory cytokines. In summary, we present a novel method to purify EGCs from rat myenteric plexus for tissue culture and to establish transformed EGC lines that retain their glial nature and functional properties. Such cell lines are now available for physiological studies of the functional properties of enteric glia in vitro.
Rühl
A
A
Department of Gastroenterology, Ruprecht-Karls-University, Heidelberg, Germany. anne.ruehl@med.uni-heidelberg.de
Trotter
J
J
Stremmel
W
W
D000818:Animals; D002460:Cell Line; D002469:Cell Separation; D017615:Enteric Nervous System / Q000166:cytology / Q000378:metabolism / Q000502:physiology*; D016762:Genes, fos / Q000235:genetics; D015151:Immunoblotting; D007150:Immunohistochemistry; D009197:Myenteric Plexus / Q000166:cytology / Q000378:metabolism / Q000502:physiology*; D009419:Nerve Tissue Proteins / Q000096:biosynthesis; D009457:Neuroglia / Q000378:metabolism / Q000502:physiology* / Q000648:ultrastructure; D012333:RNA, Messenger / Q000096:biosynthesis / Q000302:isolation & purification; D051381:Rats; D012190:Retroviridae / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction
D009419:Nerve Tissue Proteins; D012333:RNA, Messenger