This chapter describes methods used in the production of extracellular matrix (ECM), dispersion of anterior pituitary tissue, separation of dispersed cells, and development of a serum-free medium for maintenance of cultured cells. The chapter presents a series of cell dispersion techniques for the anterior pituitary in a variety of species. The chapter focuses on obtaining preparations that were acutely functional rather than on obtaining high yields of cells. For this reason, avoid using proteolytic enzymes, since the use of Viokase destroys cell surface receptors for dopamine in bovine anterior pituitary cells. Dispersed anterior pituitary cells are separated from debris and red blood cells by centrifugation through Ficoll-Hypaque as previously described for the isolation of mouse lymphocytes. The Ficoll-Hypaque is the method of choice based on ease of preparation, cell recovery, and loading capacity.
ASJC Scopus subject areas
- Molecular Biology