Klinefelter G R, Amann R P
Int J Androl. 1980 Jun;3(3):287-300. doi: 10.1111/j.1365-2605.1980.tb00119.x.
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 x 10(8)) 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. Prinicipal 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 fraction contained an average (N=7) of 23 x 10(6) 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 x 10(6) 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.
开发了一种技术，用于无菌制备富含附睾头主细胞或基底细胞的悬浮液，其数量足以进行生理学研究。为了提供无精子组织，成年大鼠的输出小管在使用前6天进行结扎。将附睾头的组织切碎并进行酶解。将解离的细胞（1.0 - 1.4×10⁸）以20毫升/分钟的缓冲液流速引入以1600转/分钟运行的贝克曼淘析转子中。除主细胞外的其他细胞通过淘析室。通过降低转子速度同时将缓冲液流速增加到30毫升/分钟来收集主细胞。最初通过转子的细胞以10毫升/分钟的流速重新引入；只有基底细胞被淘析出来。主细胞部分平均（N = 7）含有23×10⁶个细胞，其中71%为主细胞，16%为基底细胞。该部分细胞的能荷为0.92，71%的细胞排斥赤藓红-B染料。基底细胞部分通常含有30×10⁶个细胞，其中80%为基底细胞，7%为主细胞。这些细胞的能荷为0.83，85%的细胞排斥染料。
