Development of a new culture system for human lymphokine-activated killer cells

Abstract

We have developed a new culture system (termed the JCC device) for the generation and expansion of human lymphokine-activated killer (LAK) cells. The JCC device was essentially based on dialysis culture. A dialysis membrane divided the culture vessel from the outside circulation of basal medium. Human lymph node lymphocytes were cultured in the culture vessel using a culture of 100 ml at an initial cell density of 1 × 10 6/ml in the presence of recombinant human interleukin-2. The culture vessel was rotated for the proper dispersion of cells. The cell density reached up to 1.2 × 10 7/ml on day 17 of culture. A modified JCC device supplying oxygen through a silicon tube prevented oxygen depletion in the culture vessel. With this modified JCC device cell densities up to 4.3 × 10 7/ml were achieved without reduction of cytolytic activity. Cell viability was more than 90% throughout the culture. The use of this JCC device permitted the culturing of LAK cells at a high cell density. The procedure is relatively cheap, simple and less time consuming than previous methods.