Separation of human urinary proximal tubular cells from familial hypercholesterolemic homozygotes by Ficoll gradient centrifugation. Morphological and biochemical characteristics.
Separation of human urinary proximal tubular cells from familial hypercholesterolemic homozygotes by Ficoll gradient centrifugation. Morphological and biochemical characteristics.
复制标题
通过聚蔗糖梯度离心从家族性高胆固醇血症纯合子中分离人尿近端肾小管细胞。
DOI：
10.1007/bf02889879
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发表时间：
1984
期刊：
影响因子：
--
通讯作者：
KwiterovichJr,PO
中科院分区：
文献类型：
--
作者：
Chatterjee,S;Gupta,P;KwiterovichJr,PO
研究了不连续菲科尔梯度离心从家族性高胆固醇血症（FH）纯合子新鲜尿沉积物（UrS）中分离肾近端小管（PT）细胞的方法。为了比较，我们还研究了从正常人类尿路或肾脏中提取的培养细胞。未分离的PT细胞有核，90-99%的细胞不含台盼蓝。未分离和分离培养的正常PT细胞均含有大量空细胞质囊泡。相比之下，从FH纯合子的UrS中提取的类似制剂含有膜封闭的细胞质囊泡，可以用Papanicolaou （Pap）试剂染色，并且荧光素标记的抗乳糖神经酰胺抗体呈强阳性。FH纯合子的PT细胞γ-谷氨酰基转移酶和碱性磷酸酶活性分别比未分离的UrS细胞高2.0 ~ 2.5倍。我们的结论是，人类PT细胞可以通过Ficoll梯度离心从其他UrS细胞中分离出来，并且FH纯合子UrS中存在的大部分（如果不是全部的话）LacCer与PT细胞相关。纯化的PT细胞将为FH中LacCer积累的生化机制提供一个有用的模型。
The isolation of renal proximal tubular (PT) cells from the fresh urinary sediment (UrS) of familial hypercholesterolemic (FH) homozygotes using discontinuous Ficoll gradient centrifugation was studied. For comparative purposes, cultured cells derived from normal human UrS or kidney were also studied. Unfractionated PT cells were nucleated and 90–99% of the cells excluded trypan blue. Both the unfractionated and fractionated cultured normal PT cells contained numerous empty cytoplasmic vesicles. In contrast, similar preparations from the UrS of FH homozygotes contained membrane-enclosed cytoplasmic vesicles that stained with the Papanicolaou (Pap) reagent and were strongly positive with a fluorescein-labeled antibody against lactosylceramide. The PT cells of FH homozygotes contained 2.0 to 2.5 fold higher activity of γ-glutamyltransferase and alkaline phosphatase, respectively, than the unfractionated UrS cells. We conclude that human PT cells can be separated from other UrS cells by Ficoll gradient centrifugation, and that most, if not all of the LacCer present in the UrS of FH homozygotes is associated with the PT cells. Purified PT cells should provide a useful model to test the biochemical mechanisms of LacCer accumulation in FH.