Flow cytometric immunodissection of the human nephron in vivo and in vitro

In the present article, we show that flow cytometric immunodissection of ce lls immediately following their preparation from a tumor nephrectomy specim en is an accurate way of obtaining pure human primary cultures of proximal convoluted tubule origin, proximal straight tubule origin, distal tubular o rigin and/or collecting duct origin. By studying the expression of a panel of cell surface markers in these purified cultures, we could identify a num ber of markers that retain their lineage specificity in vitro. Using these appropriate stable markers, flow cytometry provides a simple yet accurate w ay of determining cell composition in previously unsorted (mixed type) tubu lar epithelial cultures in terms of proximal versus distal tubule/collectin g duct subpopulations. Both subpopulations in mixed type cultures are shown to retain functional characteristics of their in vivo counterparts (glucos e uptake, hormonal stimulation of adenylate cyclase) as well as cell type-s pecific response patterns (such as inducibility of cell adhesion and histoc ompatibility molecules), indicating the usefulness of studying cell respons es in vitro in a cell-type-dependent way. Finally we illustrate that multi- parameter flow cytometry is a powerful tool for assessing constitutive char acteristics of and/or responses by the distinct cell subpopulations present in mixed type cultures. Copyright (C) 1999 S. Karger AG, Basel.