Kreisberg J I, Hoover R L, Karnovsky M J
Kidney Int. 1978 Jul;14(1):21-30. doi: 10.1038/ki.1978.86.
Rat glomeruli were isolated by a graded sieving technique, and the nature and purity of the preparation were examined by scanning electron microscopy (SEM). A great majority of the glomeruli (86.0 +/- 6.0%) were not encapsulated, and there was very little contamination of the preparations with tubular fragments. By supplementing tissue culture medium with conditioned medium (CM) and insulin, we were able to grow single cells from dissociated rat glomeruli. From these single cells, we were able to clone and maintain in culture three distinct cell types. Also, with this specialized medium, we were able to clone these three cell types from outgrowths of whole golmeruli. One of these cell types was characterized as glomerular epithelial cells (GEC). GEC, as opposed to the other two cloned cell types obtained in this study, had cilia on their surfaces and also possessed receptors for complement (C3) in vitro. Low concentrations of the aminonucleoside of puromycin (AMNS), which have been shown to be specifically cytotoxic to GEC in vivo, were found to be cytotoxic to GEC but not to the other two glomerular cells in vitro. In addition, GEC did not contain antihemophilic factor (factor VIII), a marker for endothelium, nor were they able to phagocytose polystyrene spherules in vitro, as can mesangial cells in short-term culture.
通过分级筛分技术分离大鼠肾小球，并通过扫描电子显微镜（SEM）检查制备物的性质和纯度。绝大多数肾小球（86.0±6.0%）未被包囊，制备物中肾小管片段的污染极少。通过向组织培养基中添加条件培养基（CM）和胰岛素，我们能够从解离的大鼠肾小球中培养出单细胞。从这些单细胞中，我们能够克隆并在培养中维持三种不同的细胞类型。此外，使用这种特殊培养基，我们能够从整个肾小球的生长物中克隆出这三种细胞类型。其中一种细胞类型被鉴定为肾小球上皮细胞（GEC）。与本研究中获得的其他两种克隆细胞类型不同，GEC表面有纤毛，并且在体外也具有补体（C3）受体。低浓度的嘌呤霉素氨基核苷（AMNS）在体内已被证明对GEC具有特异性细胞毒性，在体外也被发现对GEC具有细胞毒性，但对其他两种肾小球细胞无毒性。此外，GEC不含有抗血友病因子（因子VIII），这是内皮细胞的标志物，并且它们在体外也不能像短期培养的系膜细胞那样吞噬聚苯乙烯微球。
