Preparation and properties of isolated epithelial intestinal cells from chicken cecum and jejunum.
The isolating agents, one enzymatic (hyaluronidase) and two chemical (sodium citrate and EDTA) have been used to search for the best technique to prepare suspensions of viable cells from chicken cecum and jejunum. Viability of enterocytes was assessed in terms of cell membrane integrity (trypan blue exclusion test), metabolic activity (oxygen uptake, lactate production and ATP content) and monosaccharide cumulative capacity. Results show that: In both cecum and jejunum, membrane integrity is better in cells harvested with citrate than those isolated with hyaluronidase or EDTA; The best metabolic status was found in cecal cells isolated with citrate and in jejunal cells obtained with hyaluronidase; The capacity to support alpha-methyl-D-glucoside gradients is highest in the cells harvested with citrate. The citrate-containing isolation medium is thus considered to yield epithelial cell suspensions with the best functional conditions.
D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D002432:Cecum / Q000166:cytology / Q000378:metabolism*; D002469:Cell Separation / Q000379:methods*; D002470:Cell Survival; D002645:Chickens; D006019:Glycolysis; D007583:Jejunum / Q000166:cytology / Q000378:metabolism*; D008297:Male; D010101:Oxygen Consumption
D000255:Adenosine Triphosphate