<p>Peripheral blood mononuclear cells-derived immune cells are widely used in clinical practice, in particular for cancer therapy. The growth conditions are critical for the composition and functional activity of cells. Mononuclear cells were cultured for 15 days under different conditions: in the presence of K562 cell feeder expressing 4-1BBL and mbIL-21I, or without feeder, but in the presence of L-21 or with different combinations of cytokines (IL-2, IL-15, IL-21, and IL-12) without the feeder and the cytokine profile (48 cytokines) of the culture medium in the middle (day 7) and at the end (day 14) of culturing was compared. In the presence of the feeder, the cells produced IFNγ, RANTES, MIP-1α, IL-9, IL-6, and CF-CSF in high concentrations (&gt; 5000 pg/ml) and less intensively produced MDC; NK cells predominated in the culture. These cytokines promote proliferation of immune cells, participate in antitumor response (IFNγ and RANTES) and have a dual effect on the growth of some tumor cell lines (IL-6, IL-9, and VEGF). The conditioned medium, especially the medium from mononuclears grown without feeder, inhibited the growth of lung cancer cell line A549. The studied methods for mononuclear cell culturing increase the functional and antitumor activities of NK cells via stimulation of cytokine production.</p>

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Secretome and Cellular Composition of Peripheral Blood Mononuclear Cells Cultured in the Presence of Cell Feeder or Interleukins

  • J. M. Rozenberg,
  • R. V. Zhurikov,
  • A. S. Oleinik,
  • S. A. Roumiantsev,
  • E. V. Abakushina

摘要

Peripheral blood mononuclear cells-derived immune cells are widely used in clinical practice, in particular for cancer therapy. The growth conditions are critical for the composition and functional activity of cells. Mononuclear cells were cultured for 15 days under different conditions: in the presence of K562 cell feeder expressing 4-1BBL and mbIL-21I, or without feeder, but in the presence of L-21 or with different combinations of cytokines (IL-2, IL-15, IL-21, and IL-12) without the feeder and the cytokine profile (48 cytokines) of the culture medium in the middle (day 7) and at the end (day 14) of culturing was compared. In the presence of the feeder, the cells produced IFNγ, RANTES, MIP-1α, IL-9, IL-6, and CF-CSF in high concentrations (> 5000 pg/ml) and less intensively produced MDC; NK cells predominated in the culture. These cytokines promote proliferation of immune cells, participate in antitumor response (IFNγ and RANTES) and have a dual effect on the growth of some tumor cell lines (IL-6, IL-9, and VEGF). The conditioned medium, especially the medium from mononuclears grown without feeder, inhibited the growth of lung cancer cell line A549. The studied methods for mononuclear cell culturing increase the functional and antitumor activities of NK cells via stimulation of cytokine production.