Isolation of Principal Cells and Basal Cells by Elutriation of Suspensions of Rat Epididymal Tissue

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Abstract
A technique was developed to aseptically prepare suspensions enriched in principal cells or basal cells of the caput epididymidis in number sufficient for physiological studies. To provide sperm-free tissue the efferent ducts of adult rats were ligated 6 days before use. Tissue from the caput epididymidis was minced and dissociated enzymatically. Dissociated cells (1.0-1.4 .times. 108) were introduced with a buffer flow of 20 ml/min into a Beckman elutriator rotor operating at 1600 rpm. Cells other than principal cells passed through the elutriation chamber. Principal cells were collected by decreasing rotor speed while increasing buffer flow to 30 ml/min. Cells initially passing through the rotor were reintroduced at a flow of 10 ml/min; only basal cells were elutriated. The principal-cell fractions contained an average (N = 7) of 23 .times. 106 cells of which 71% were principal cells and 16% were basal cells. Cells in this fraction had an energy charge of 0.92 and 71% excluded erythrosin-B dye. The basal-cell fraction typically contained 30 .times. 106 cells of which 80% were basal cells and 7% were principal cells. These cells had an energy charge of 0.83 and 85% excluded dye.