Lymphokine-activated killer cell generation and recovery. Comparison of an automated cell processing device and a manual procedure.

Abstract

This report describes the separation or processing of leukapheresis-derived peripheral blood mononuclear cells, their culture in the presence of interleukin-2 (IL-2) to generate lymphokine-activated killer (LAK) cells, and the harvest of LAK cells using the Du Pont SteriCell processor and culture containers. The report compares this automated closed system to the standard National Cancer Institute (NCI) manual protocols. Lymphocytes (approximately 2.0 x 10(10)) were harvested on the Cobe 2997 blood cell separator for each set of experiments. A set of experiments using automated Ficoll-Isopaque (FI) separation and a set of experiments using an automated wash for platelet reduction were compared to the respective manual methods. SteriCell culture containers were compared to roller bottles, and automated harvest was compared to manual harvest. The cytolytic activity of LAK cells was assayed against that of Daudi cells in a standard 51Cr release assay. The cytolytic activity of LAK cells prepared by a manual method and by the automated SteriCell method are equivalent. The manual and SteriCell FI separation procedures were substantially equivalent. The SteriCell harvest method gave higher recoveries than the manual harvest method (p = 0.04 and p = 0.07), but the overall recovery of LAK cells was not significantly different. Differential counts on the products were similar. Platelet reduction by the SteriCell automated wash procedure was greater than that by the manual procedure (p = 0.05). The SteriCell system represents an acceptable alternative to the manual methods for LAK cell generation and recovery.