Autologous lymphocytes prevent the death of monocytes in culture and promote, as do GM-CSF, IL-3 and M-CSF, their differentiation into macrophages

• Published in: Journal of immunological methods Vol. 159; no. 1-2; p. 29
• Main Authors: Lopez, M, Martinache, C, Canepa, S, Chokri, M, Scotto, F, Bartholeyns, J
• Format: Journal Article
• Language: English
• Published: Netherlands 26-02-1993
• Subjects: CD3 Complex - analysis; Cell Differentiation - drug effects; Cell Survival; Cells, Cultured; Culture Media, Conditioned; Granulocyte-Macrophage Colony-Stimulating Factor - pharmacology; Humans; Interleukin-3 - pharmacology; Lymphocytes - physiology; Macrophage Colony-Stimulating Factor - pharmacology; Macrophages - drug effects; Macrophages - physiology; Monocytes - physiology
• ISSN: 0022-1759
• DOI: 10.1016/0022-1759(93)90138-W

Blood monocytes collected by apheresis from healthy donors were differentiated in vitro to macrophages which were subsequently activated with recombinant human interferon-gamma. 7 day cultures were established by seeding Ficoll-separated mononuclear cells or elutriation-purified monocytes under different culture conditions. The best macrophage yields required the seeding of mononuclear cells (instead of purified monocytes) in teflon bags with a high air-liquid surface interface. The effects of GM-CSF, IL-3 and M-CSF on the macrophage yield were then evaluated. GM-CSF increased the average yield by 3.6- and 2.3-fold when purified monocytes or total mononuclear cells were seeded respectively. The corresponding increases with IL-3 were 2.5- and 2.1-fold respectively and with M-CSF 1.2- and 1.4-fold respectively. Macrophages matured under these various conditions displayed similar CD14, CD64, CD71, HLA-DR and Max 1 antigen expression and similar in vitro anti-tumoral activity against U937 cells. Culturing in the presence of cytokines permits the large scale production of activated macrophages for adoptive immunotherapy trials.