Incorporation of 3 H-Thymidine into PHA-Stimulated Rabbit Peripheral Blood Lymphocytes. Kinetics of the Response 1

Abstract

Summary

Rabbit PBL were shown to incorporate predictably high levels of ³H-TdR following stimulation with PHA. The conditions necessary for optimal stimulation of rabbit PBL were: (a) separation of lymphocytes from whole blood by the Hypaque-Ficoll technique; (b) 2 × 10⁶ PBL per culture in RPMI-1640 containing 5–10% heat-inactivated, autologous plasma; (c) 48 hr culture period; (d) 5 μg/ml PHA-P; and (e) 1 μCi/ml ³H-TdR added to cultures 22 hr before termination. Under these conditions, individual and group variations in the response to PHA were slight, and PBL responsiveness to other mitogens as well as to antigen were good.