Enoch P G, Hemstreet G P, Fine P R
Invest Urol. 1981 Mar;18(5):345-8.
Five normal human kidneys were disaggregated with three combinations of enzymes. The combination of collagenase 0.5 mg per ml of 11 additional kidneys. The viability of cells in suspension ranged from 75 to 94 per cent with a mean viability of 85.3 +/- 1.6 per cent. Disaggregated kidney cells were fractionated in isopyknic and isokinetic gradients. After isokinetic sedimentation, kidney tubule cells were predominantly in fraction 22 +/- 1 whereas lymphocytes were consistently located in fraction 8 +/- 1. Kidney epithelial cells were enriched to a 90.4 +/- 2.2 per cent (79.0 to 96.0 per cent) purity by isokinetic gradient centrifugation. Controlled rate freezing of cells provided material for repeated experiments, and short-term tissue culture before cell separation increased the proportion of viable cells in suspension.
用三种酶组合对五个正常人类肾脏进行解离。每毫升含0.5毫克胶原酶的组合用于另外11个肾脏。悬浮细胞的活力在75%至94%之间,平均活力为85.3±1.6%。将解离的肾细胞在等密度和等速梯度中进行分级分离。等速沉降后,肾小管细胞主要在22±1级分中,而淋巴细胞始终位于8±1级分中。通过等速梯度离心,肾上皮细胞纯度提高到90.4±2.2%(79.0%至96.0%)。细胞的控制速率冷冻为重复实验提供了材料,细胞分离前的短期组织培养增加了悬浮中活细胞的比例。