Helms S R, Brazeal F I, Bueschen A J, Pretlow T G
Am J Pathol. 1975 Jul;80(1):79-90.
Collagenase, trypsin, and pronase were used in separate, parallel experiments to obtain cell suspensions from human prostates obtained from surgical resections and autopsies. All of the examined prostates demonstrated benign hyperplasia. The dissociation using pronase gave both the largest number of nucleated cells and the largest proportion of viable cells from prostates. Surgically resected prostates gave a larger number of cells per gram of tissue than prostates obtained from autopsy. Cells obtained from surgically resected prostates were separated both by isopycnic sedimentation and by velocity sedimentation in a previously described isokinetic gradient. We studied fifteen prostates obtained at surgery; using pronase, we obtained 2.1 plus or minus 3.5 times 10(6) cells/g. Of these cells, 34.0 plus or minus 14.7% contained histochemically demonstrable acid phosphatase. Cells from six prostates were separated by velocity sedimentation in the isokinetic gradients; in the purest fractions from the isokinetic gradients, 81.0 plus or minus 12.2% of nucleated cells had histochemically demonstrable acid phosphatase. The cells in the purest fractions appeared to be epithelial cells. More than 99% of these separated cells excluded trypan blue. Isopycnic sedimentation was not an effective means of purifying epithelial cells from human prostates.
在单独的平行实验中使用胶原酶、胰蛋白酶和链霉蛋白酶,从手术切除和尸检获得的人类前列腺中获取细胞悬液。所有检查的前列腺均显示为良性增生。使用链霉蛋白酶进行解离时,从前列腺中获得的有核细胞数量最多,活细胞比例也最高。手术切除的前列腺每克组织产生的细胞数量比尸检获得的前列腺更多。通过等密度沉降和在先前描述的等速梯度中进行速度沉降,对手术切除的前列腺获得的细胞进行分离。我们研究了15个手术获取的前列腺;使用链霉蛋白酶,我们获得了2.1±3.5×10⁶个细胞/克。在这些细胞中,34.0±14.7%含有组织化学可显示的酸性磷酸酶。对来自6个前列腺的细胞在等速梯度中进行速度沉降分离;在等速梯度最纯的组分中,81.0±12.2%的有核细胞含有组织化学可显示的酸性磷酸酶。最纯组分中的细胞似乎是上皮细胞。这些分离的细胞中超过99%排斥台盼蓝。等密度沉降不是从人类前列腺中纯化上皮细胞的有效方法。