Improved culture conditions for quantitative evaluation of interleukin 2 production by frozen human lymphocytes.
Several culture parameters were studied in order to establish methods for optimal and reproducible production of interleukin 2 (IL2) by thawed lymphocytes. Standard conditions, considered optimal for production by freshly separated lymphocytes (culture medium RPMI 1640 + 1% normal human serum + 10 micrograms/ml PHA), gave low and poorly reproducible results. An increased concentration of human serum (10 and 20%) in the medium improved production but best results were obtained by adding polyethylene glycol (PEG 6000, 0.1 mg/ml) to the culture medium. Furthermore, with the addition of PEG 6000, results became highly reproducible, thus permitting valid comparison of in vitro IL2 production by lymphocytes from normal donors and patients.
D001769:Blood; D002478:Cells, Cultured; D003470:Culture Media; D005615:Freezing; D006801:Humans; D007376:Interleukin-2 / Q000235:genetics* / Q000302:isolation & purification; D007700:Kinetics; D008213:Lymphocyte Activation; D008214:Lymphocytes / Q000276:immunology / Q000502:physiology*; D008297:Male
D003470:Culture Media; D007376:Interleukin-2