Chapter 89 - Current Methodologic Issues in Cell and Tissue Culture

Abstract

Publisher Summary This chapter focuses on the various in vitro methods used in skeletal biology. Bone organ (tissue) cultures have been in use since the pioneering studies of Strangeways and Fell in the 1920s, while the isolation and culture of bone cells came into practice nearly half a century later. Most of the in vitro culture systems used in skeletal biology today are remarkably similar in principle and procedure to their early ancestors. The day-to-day techniques of tissue isolation, feeding, subculturing, cloning, and preservation are much the same as they were decades ago, with the exception that most materials are now made of disposable plasticware rather than glass, and most media are purchased ready to use rather than prepared from scratch. Organ cultures offer investigators the opportunities to study a tissue system with much of its architectural hierarchy intact, but without interference from the rest of the animal and its variable systemic influences. Bone organ culture systems have been used primarily to study dynamic events and their regulation, which include bone formation and matrix synthesis, bone resorption, and tissue morphogenesis. All of these processes are associated with tissue growth and development, and thus are amenable to study using embryonic, fetal, or newborn tissues that have high metabolic activities and can be maintained easily over short periods in organ culture. Cell cultures offer a distinctly different approach to in vitro biology than organ cultures. The cell culture approach treats mammalian cells virtually like bacterial cultures, which allows the investigator to study the cells' intrinsic functions like proliferation, differentiation, adhesion, migration, matrix synthesis, and death, as well as their interactions with each other and with their environment.