This chapter discusses techniques for the production and maintenance of human T cell clones, their growth characteristics and longevity in vitro, culture age-associated changes of parameters such as telomere lengths, DNA damage, and so on. Cloning peripheral T cells from young healthy donors can be performed with a high cloning efficiency (CE), suggesting that these particular cloning conditions are favorable for outgrowth of a majority of T cells. The CE was equally high when the cells were derived from a patient with chronic myelogenous leukemia, a donor putatively subjected to a high level of antigenic stress. CE was also similar in the extra-thymic T cell differentiation cultures employed. According to the protocol for cloning, for ease of handling and as a precaution against contamination, stack plates and aluminum foil should be used. Incubate for about a week and then examine the plates using an inverted microscope. T cell functions are mediated by intracellular signaling via surface receptors such as the antigen receptor and the costimulator CD28 from the membrane through the cytoplasm to the nucleus.
ASJC Scopus subject areas
- Biochemistry, Genetics and Molecular Biology(all)