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Collagenase-dispersed cells from the guinea-pig ileum were prepared and Ca2+ release into Ca2+-depleted solution from the isolated single cells obtained by the centrifugation of the dispersed cells on isotonic sucrose solution was determined with a Ca2+-selective electrode. A technique employing an isotonic sucrose solution for washing isolated cells permitted removal of contaminating extracellular fluids and obtaining the isolated cells with minimum loss of cellular Ca2+. The release of Ca2+ from the dispersed cells consisted of at least two phases. The Ca2+ release into an isotonic sucrose-Tris solution (ISTS) was significantly reduced and the early phase of the Ca2+ release disappeared. At 16 degrees C, Ca2+ release depended on the composition of the bathing solution in a Ca2+-free salt solution, the later phase of Ca2+-release was abolished whereas in ISTS both phases disappeared. Furthermore, Ca2+ release was significantly reduced after the treatment of the dispersed cells with disperse (1,500 units/ml) for 10 min. These results show that Ca2+ release into Ca2+-depleted solution from the isolated ileal cells with minimum loss of cellular Ca2+ show a biphasic curve and suggest that Ca2+ sources responsible for these phases may be of distinct origin.

Determination of Ca2+-release from the isolated smooth muscle cells in suspension from the guinea-pig ileum.