|Dombrink Kurtzman, Mary Ann|
Submitted to: Advances in Peritoneal Dialysis
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 4/22/1998
Publication Date: N/A
Citation: Interpretive Summary: The study was undertaken to compare extraction techniques which will be used to gain an understanding of the sphingoid base (fat components) composition in the mesothelium (cells in the peritoneum, which lines the cavity of the abdomen) of individuals undergoing peritoneal dialysis. People undergoing this procedure have impaired kidney function and may represent a potentially vulnerable population to exposure to chronic low doses of fungal toxins. We examined human mesothelial cells grown in culture to determine the amounts of the sphingoid bases, sphingosine, and sphinganine. Two different extraction methods were used. A difference was observed based on the method used to digest the cells. This technical information will aid future research on the impact that environmental exposure to microbial toxins has on the fat composition in mesothelial cells.
Technical Abstract: Sphingolipids are emerging as important regulators of mammalian cell biology. In this study, six separate preparations of human omental mesothelial cells in vitro were used to examine for the content of free sphingosine and sphinganine and the total levels of these sphingoid bases in ceramide-containing sphingolipids. Two HPLC methods for determination of sphingoid base levels in cultured cells were compared. The rapid HPLC method was found to yield the highest recovery of internal standard. Mesothelial cells initially isolated by collagenase digestion of the omentum were found to have higher free and total sphingoid base levels than cells isolated by trypsin-EDTA digestion. Use of sphingoid base levels to gain insights into the status of cellular nutrition, inflammation, programmed cells death, exposure to microbial toxins, cytokines and growth factors within the peritoneum will require a systematic description of sphingolipids in normal, diseased, and dialyzed mesothelium.