Development of a new culture system for human lymphokine-activated killer cells.
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 volume of 100 ml at an initial cell density of 1 X 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 X 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 X 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.
Murata
M
M
Department of Immunology, Kyushu University, Fukuoka, Japan.
Yano
T
T
Togami
M
M
Yasumoto
K
K
Sugimachi
K
K
Kimura
G
G
Nomoto
K
K
D002452:Cell Count; D002478:Cells, Cultured; D003470:Culture Media; D003601:Cytotoxicity Tests, Immunologic; D003956:Dialysis; D004867:Equipment Design; D005947:Glucose / Q000378:metabolism; D006801:Humans; D007376:Interleukin-2 / Q000494:pharmacology; D015979:Killer Cells, Lymphokine-Activated / Q000276:immunology / Q000378:metabolism; D007773:Lactates / Q000378:metabolism; D019344:Lactic Acid; D008198:Lymph Nodes / Q000166:cytology; D008214:Lymphocytes / Q000276:immunology / Q000378:metabolism; D008566:Membranes; D008722:Methods; D010100:Oxygen / Q000494:pharmacology; D011994:Recombinant Proteins
D003470:Culture Media; D007376:Interleukin-2; D007773:Lactates; D011994:Recombinant Proteins; D019344:Lactic Acid; D005947:Glucose; D010100:Oxygen