A simple and inexpensive procedure is described which allows reproducibly the isolation of rat pancreatic acinar cells. Using only small quantities of commercially available collagenase without addition of any further protease, a cell population consisting of about 95% of acinar cells can be obtained within about 95 min. Cell yield is 40% as calculated on a dry weight basis. Enzyme activities measured within final suspensions of isolated cells are: amylase, 1.17 +/- 0.27 amylase units.(mg d.w.)-1; lipase, 23.78 +/- 6.02 nkat.(mg d.w.)-1 and alanine aminotransferase, 0.895 +/- 0.236 nkat.(mg d.w.)-1. Isolated cells are morphologically intact as seen by electron microscopy and retain their viability for more than 3 h, even when incubated at 37 degrees C without any substrate and protease inhibitor, as revealed by their ability to exclude trypen blue. Therefore, acinar cells isolated in this manner may prove useful for investigations at cellular level into pathogenetic mechanisms underlying pancreatic diseases.