Abstract
Functionally and anatomically intact acinar cells were obtained from the parotid gland of the rat by enzymatic dispersion. After exsanguination of the animal, the duct system of the gland was filled with a solution of hyaluronidase and collagenase (0.1%) in Ca±Mg free Hank's salt medium. The parotid was removed, minced and the tissue was incubated in the same enzyme solution for 60 min at 37°C. Then, the suspension was filtered through nylon mesh, the clumps of cells were separated by centrifugation, were suspended in a solution of trypsin (0.1%) and reincubated for 20 min. Finally, the cells were dispersed by pipetting the suspension 10 times, washed and then dispersed by pipetting the suspension 10 times, washed and then suspended in balanced Hank's solution containing bovine serum albumin (2 gm/100 ml). Yield of cells was ≃60% of the original gland tissue. The cells were anatomically intact and did not stain with 0.2% trypan blue for up to 8 hours. They showed normal respiration (O2 uptake: 21.4 ± 2.7 μl/hours·mg protein) and responded to stimulants of sectory activity (epinephrine, isoproterenol, dibutyryl cyclic. AMP, theophylline and NaF) by secreting amylase into the medim at rates approximately equal to those observed in the intact gland in situ.
This method permits the investigation of aspects of the secretory processes of exocrine glands which cannot be studied in the intact glands of experimental animals in situ or in gland slices. Furthermore, as preliminary experimetns have shown, living acinar cells for similar secretory studies can be obtained from human salivary glands and pancreas immediately after death.
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Mangos, J., Mcsherry, N. Secretory function of isolated parotid acinar cells. Pediatr Res 5, 418 (1971). https://doi.org/10.1203/00006450-197108000-00196
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DOI: https://doi.org/10.1203/00006450-197108000-00196