| Popis: |
In this chapter we will briefly review the techniques available for isolation and culture of type II pneumocytes and describe in detail a technique for the rapid isolation of adult rat type II cells. The lung consists of 40 different cell types of which type II cells make up 15% [1]. Type II alveolar epithelial cells are important for normal lung function [2] and in regeneration of lung epithelium following injury [3, 4]. They synthesize and secrete surfactant phospholipids and proteins that are required for lowering surface tension at low lung volumes. Type II cells also participate in many other functions such as inflammatory and immune responses, oxidant metabolism, fluid transport, and electrolyte transport. To investigate how these functions are regulated, it is important to study pure type II cell populations. Many different methods have been developed to isolate type II cells for study in vitro (reviewed in [5]). In vitro studies are still limited by the rapid loss of differentiated functions of type II cells in culture [6]. Furthermore, the most commonly used isolation techniques require culturing type II cells for 1–2 days before they are available for study. Most techniques that permit isolation and immediate study of type II cells either have limited cell yields [5] or require specialized equipment not routinely available. In this chapter, we will discuss the techniques available for type II cell isolation and describe a method for isolation of adult rat type pneumocytes for immediate use. |
