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

Citation
Mjf. Helbert et al., Flow cytometric immunodissection of the human nephron in vivo and in vitro, EXP NEPHROL, 7(5-6), 1999, pp. 360-376
Citations number
45
Categorie Soggetti
Urology & Nephrology","da verificare
Journal title
EXPERIMENTAL NEPHROLOGY
ISSN journal
10187782 → ACNP
Volume
7
Issue
5-6
Year of publication
1999
Pages
360 - 376
Database
ISI
SICI code
1018-7782(199909/12)7:5-6<360:FCIOTH>2.0.ZU;2-5
Abstract
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.