The use of hyperosmolar, intracellular-like solutions for the isolation of epithelial cells from guinea-pig small intestine.
Isolated small intestine epithelial cells were prepared by using either (a) hyperosmolar, low sodium, high potassium containing (intracellular-like) solutions, or (b) isosmolar, high sodium, low potassium containing (extracellular-like) solutions. Both (a) and (b) cells show high viability as estimated by Trypan blue exclusion, oxygen consumption, cellular ATP content, lactate-dehydrogenase liberation, intracellular ion concentrations and significant Na+-dependent alanine and uridine uptakes. Although (a) and (b) cells show in the cold similar ion concentration, after reincubation at 37 degrees C for 30 min (a) cells show intracellular ion concentrations of 31 mM Na, 129 mM K and 88 mM Cl, whilst (b) cells have 71 mM Na, 93 mM K and 102 mM Cl. Cells prepared with (a) concentrate much more alanine and uridine than cells prepared with (b), probably because the latter have a lower Na+ gradient across the plasma membrane. Cells prepared with intracellular-like solutions would be an ideal system to study Na+-dependent transport mechanisms and the regulatory systems of intracellular ion concentrations.
D000255:Adenosine Triphosphate / Q000032:analysis; D000818:Animals; D002469:Cell Separation / Q000379:methods; D003080:Cold Temperature; D004847:Epithelial Cells; D006168:Guinea Pigs; D007421:Intestine, Small / Q000166:cytology*; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008297:Male; D008854:Microscopy, Electron; D009994:Osmolar Concentration; D010101:Oxygen Consumption; D011188:Potassium; D012964:Sodium; D012996:Solutions
D012996:Solutions; D000255:Adenosine Triphosphate; D012964:Sodium; D007770:L-Lactate Dehydrogenase; D011188:Potassium