ABSTRACT
Protocol 2.4: Incorporation of [3H]thymidine into acid-insoluble material
Protocol 2.5: Autoradiography of labelled nuclei 28
Protocol 2.6: BrdU incorporation and staining 29
Protocol 2.7: Bromodeoxyuridine anti-bromodeoxyuridine demonstration of biopsy samples
Protocol 2.8: Analysis of the cell cycle using flow cytometry 34
2.5 Measurement of cell proliferation in vivo 35
2.6 Tracking cell division in vitro and in vivo using CFSE dilution and flow cytometry
2.7 References 37
2.1 Introduction
Cell division and cell proliferation are fundamental processes in all living organisms. In mammals, cell proliferation and the growth factors, growth inhibitors and cytokines which regulate it, are essential during embryonic development, tissue and organ growth, and for several physiological processes in the adult state such as haematopoiesis, wound healing and pregnancy. For much of their lifespan, the differentiated cells of many adult mammalian tissues exist in a viable, non-proliferating state. Many such cells retain the capacity to proliferate and will do so during the repair of damaged tissue, or can be stimulated to reinitiate DNA synthesis and cell division when placed in culture and challenged with suitable growth factors [1]. The abnormal hyperplastic or neoplastic multiplication of normally non-proliferating adult cells in vivo also plays a central role in tumourigenesis, atherosclerosis and other disease processes. Consequently, techniques for the accurate, reliable and rapid evaluation of cell proliferation are among the most widespread and important in clinical and basic biological research.
