A primary culture of proximal tubule cells of the mouse kidney on collagen coated membranes: characterization, electrophysiological properties and response to stress factors

Abstract

Over the years, a large number of in vitro models ranging from renal cell lines to isolated perfused tubules have been established. However, epithelial cell lines had to undergo a number of genetic changes in order to survive indefinitely in vitro. On the other hand, the isolated perfused tubule is a complex system to study. As a consequence, primary epithelial cell culture systems are needed to check the physiological significance of studies done with established cell lines and in some cases they might be the only way to study specific tubular cell function characteristics in particular (patho)physiological and pharmacological conditions. Furthermore, the increasing use of transgenic and knock-out mouse models should be supported by an in vitro model to study the cellular phenotype. Major inherent technical advantages of cultured cells for the study of epithelial transport include their ability to be studied in a controlled environment without systemic neurons and hormonal influences, and their easy accessibility for electrophysiological and imaging techniques. However, primary cell culture also has its limitations such as incomplete differentiation of cells or a limited lifespan, among others. The objective of this thesis was to establish a primary cell culture of mouse proximal tubule cells. The primary cells were extensively characterized for their proximal tubule origin. To correlate our findings in primary cell cultures with data in the literature, a detailed description of proximal tubule cell morphology and ( electro-) physiology is given in section l. 2, l. 3 and 1.4. Finally, we used our cell cultures as a model for the study of renal tubulointerstitial fibrosis. The mechanisms involved in tubulointerstitial fibrosis are described in section 1.5. At the end of this chapter, the different aims of this thesis are summarized.