Comparison of the use of bulk to micro culture of cell preparations for lymphokine-activated cytotoxicity assays

✍ Scribed by Loraine H. Anderson; Thomas L. McDonald; Geoffrey M. Thiele; Lynell W. Klassen

• Publisher: John Wiley and Sons
• Year: 1992
• Tongue: English
• Weight: 537 KB
• Volume: 6
• Category: Article
• ISSN: 0887-8013
• DOI: 10.1002/jcla.1860060302

✦ Synopsis

Abstract

Different assay systems have been used to quantitate lymphokine‐induced natural cytotoxic activity as a measure of immune status. This study compares the effects of inducing cytotoxicity in a bulk culture system, where effector cells are transferred to a micro culture well for assay, to a micro culture system where the effector cells are not transferred. The effector/target ratio for both the bulk and micro culture systems was calculated using the number of viable effector cells present at the time of target cell addition. After over‐night incubation with interleukin‐2 (IL‐2), the lytic activity of murine spleen cells to targets using a micro culture system was increased two‐fold over the bulk culture method. This increase was amplified further after 5 days of activation with IL‐2, in that the micro culture system resulted in a four‐fold increase in cytotoxic activity. The loss of some adherent cells in the bulk culture system did not explain the overall decrease in recovered cytotoxicity. The difference appeared to be related to cell loss during centrifugation. Therefore, the E/T ratios are different in the two systems if not corrected for the number of viable cells. © 1992 Wiley‐Liss, Inc.