Culturing steroidogenic cells

This chapter discusses the culturing steroidogenic cells. Cultures of steroidogenic cells have been invaluable in many studies of the molecular biology, cell biology, and physiology of steroidogenic tissues. In this chapter the procedures for preparation, growth, and storage of steroidogenic cells from the adrenal cortex, ovary, and testis are described. Preparation of a primary cell suspension requires first the dissection of steroidogenic tissue away from other tissue components, cell dissociation using enzymes and/or mechanical dispersion, and, in some cases, separation of cell types from a mixed preparation by gravity sedimentation. In some species, abundant stored lipid droplets may make cells fragile and susceptible to mechanical damage. Tissue fragments resulting from the dissection procedure may be dissociated using crude collagenase. The enzymes in crude collagenase have a short half-life in solution because of self-digestion; therefore, collagenase solutions should not be prepared in advance or reused.