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从附睾体中分离上皮细胞,并分析其甘油磷酸胆碱、唾液酸和蛋白质。

Isolation of epithelial cells from the corpus epididymidis and analysis for glycerylphosphorylcholine, sialic acid, and protein.

作者信息

Hoffmann D S, Killian G J

出版信息

J Exp Zool. 1981 Jul;217(1):93-102. doi: 10.1002/jez.1402170110.

Abstract

Enzymatically dispersed cells from the rat corpus epididymidis were separated by unit gravity sedimentation and centrifugal elutriation. Based on differential cell counts, principal cells isolated by centrifugal elutriation were 55% pure, while basal cells and fibroblasts were 88% and 61% pure, respectively. Compared to unit gravity separation, purity was an average of 7% higher in principal and basal cell fractions obtained by elutriation. Viability was greater than 87% by either method, but yields following elutriation averaged 23% higher for basal cells, 78% higher for principal cells, and 290% for fibroblasts than corresponding yields following unit gravity separation. Analyses for epididymal secretory products indicated that cellular content (mumoles/10(9) cells) of glycerylphosphorylcholine (GPC), sialic acid, and protein was greater in principal cells than other cell types (p less than .05). When expressed on the basis of cellular volume, concentrations of GPC and protein also were significantly greater in basal and principal cells than fibroblasts (p less than .05). Caput sperm contained significantly more GPC than either corpus or cauda sperm (p less than .005). Protein concentrations in caput and corpus sperm were similar, but concentrations were lower in cauda sperm (p less than .005). Levels of sialic acid did not vary among sperm from different epididymal regions. It was concluded that principal cells from the corpus epididymidis contain major quantities of three epididymal secretory products and that regional differences in the function of the epithelium are present.

摘要

通过单位重力沉降和离心淘析法分离大鼠附睾体的酶解分散细胞。基于细胞计数差异,通过离心淘析分离得到的主细胞纯度为55%,而基底细胞和成纤维细胞的纯度分别为88%和61%。与单位重力分离法相比,通过淘析法获得的主细胞和基底细胞组分的纯度平均高出7%。两种方法的细胞活力均大于87%,但淘析法后的基底细胞产量平均比单位重力分离法高出23%,主细胞高出78%,成纤维细胞高出290%。附睾分泌产物分析表明,主细胞中甘油磷酸胆碱(GPC)、唾液酸和蛋白质的细胞含量(微摩尔/10⁹细胞)高于其他细胞类型(p<0.05)。以细胞体积为基础表示时,基底细胞和主细胞中GPC和蛋白质的浓度也显著高于成纤维细胞(p<0.05)。附睾头部精子中的GPC含量显著高于附睾体部或尾部精子(p<0.005)。附睾头部和体部精子中的蛋白质浓度相似,但尾部精子中的浓度较低(p<0.005)。不同附睾区域精子中的唾液酸水平没有差异。得出的结论是,附睾体的主细胞含有大量三种附睾分泌产物,并且上皮功能存在区域差异。

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