Cell culture techniques

Abstract

INTRODUCTION Cell culture is a technique that involves the isolation and in vitro maintenance of cells isolated from tissues or whole organs derived from animals, microbes or plants. In general, animal cells have more complex nutritional requirements and usually need more stringent conditions for growth and maintenance than microbial cells. By comparison, microbes and plants require less rigorous conditions and grow effectively with the minimum of needs. Regardless of the source of material used, practical cell culture is governed by the same general principles, requiring a sterile pure culture of cells, the need to adopt appropriate aseptic techniques and the utilisation of suitable conditions for optimal viable growth of cells. Once established, cells in culture can be exploited in many different ways. For instance, they are ideal for studying intracellular processes including protein synthesis, signal transduction mechanisms and drug metabolism. They have also been widely used to understand the mechanisms of drug actions, cell-to-cell interactions and genetics. Additionally, cell culture technology has been adopted in medicine, where genetic abnormalities can be determined by chromosomal analysis of cells derived, for example, from expectant mothers. Similarly, viral infections can be assayed both qualitatively and quantitatively on isolated cells in culture. In industry as well as academic drug discovery, cultured cells are used routinely to test both the pharmacological and toxicological effects of pharmaceutical compounds. This technology thus provides a valuable tool to scientists, offering a user-friendly system that is relatively cheap to run and the exploitation of which avoids the legal, moral and ethical questions generally associated with animal experimentation. More importantly, cell culture also presents a tremendous potential for future exploitation in disease treatment, where, for instance, defective or malfunctioning genes could be corrected in the host's own cells and transplanted back into the host to treat a disease. Furthermore, successful development of culture techniques for stem cells provides a much needed cell-based strategy for treating diseases where organ transplant is currently the only available option. In this chapter, fundamental information required for standard cell culture, together with a series of principles and protocols used routinely in growing animal and bacterial cells are discussed. Additionally, a section has been dedicated to human embryonic stem cell culture, outlining techniques that are now becoming routine for stem cell culture.