Culture technique of human pituitary adenoma cells

Cell cultures are indispensable to the study of human pituitary adenomas Nevertheless, in vitro studies of human pituitary adenomas have been hindered by the small amount of tissue available and by difficulties in establishing monolayer cultures using standard culture techniques owing to poor cell attachment to ordinary plastic flasks It is well known that pituitary adenoma cells attach poorly to plastic culture flasks, form floating aggregations in the medium, and are easily washed out by medium change (1) Pituitary adenoma cells do not grow rapidly enough for subcultures, so the primary culture is maintained for a relatively long period However, such cells are occasionally overgrown by fibroblasts, which eventually become free floating in the medium (1, 2). Plastic dishes coated with extracellular matrix have been used recently for culturing of pituitary adenoma cells (3, 4), but good results are not necessarily obtained using this method in our experience. Therefore, in this chapter, I describe simple techniques for obtaining an in vitro model system of human hormoneproducing pituitary adenoma, which has relatively firm cell attachment and well-preserved hormonal function (5). In brief, human pituitary adenoma cells are cultured on a microporous membrane (6) coated with a basement membrane extract under the control of seeding medium volume (5).