Abstract
This chapter explores periodate-induced blast transformation in mixtures of rat lymphoid cells. In a study described in the chapter, lymphoid cell populations from Lewis rats were prepared. Cells were cultured in RPMI-1640 containing 20% heat-inactivated horse serum in Microtest II plates. Mixing experiments were performed at constant total cell concentration by varying the proportions of the two cell populations over the entire range between the two pure populations. Incorporation of 3 H-dT was determined by pulse-labeling under flooding conditions with low specific activity material to minimize possible complications because of variable endogenous cold dT pools. It was verified at all time points that 3 H-dT incorporation was a linear function of cell concentration and that there was no depletion of culture nutrients. Chromosome analysis of colchicine-arrested cells was based on the procedures of Hungerford and Hungerford and Nowell. The results showed that rat lymph node cells (LNC), thoracic duct lymphocytes (TDL), and thymocytes each responded optimally after treatment with 1.2 mM periodate and exhibited identical kinetics with a peak of 3 H-dT incorporation between 48 and 54 h. However, LNC incorporated 10–20 times more 3 H-dT than TDL or thymocytes.
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