Lopez M, Martinache C, Canepa S, Chokri M, Scotto F, Bartholeyns J
Inserm U76, Centre National de Transfusion Sanguine, Etablissement Saint-Antoine, Paris, France.
J Immunol Methods. 1993 Feb 26;159(1-2):29-38. 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.
通过单采术从健康供体采集的血液单核细胞在体外分化为巨噬细胞，随后用重组人干扰素-γ激活。通过在不同培养条件下接种经Ficoll分离的单核细胞或淘洗纯化的单核细胞建立7天培养物。获得最佳巨噬细胞产量需要在具有高气液表面界面的聚四氟乙烯袋中接种单核细胞（而非纯化的单核细胞）。然后评估GM-CSF、IL-3和M-CSF对巨噬细胞产量的影响。当分别接种纯化的单核细胞或总单核细胞时，GM-CSF使平均产量分别提高了3.6倍和2.3倍。IL-3的相应提高分别为2.5倍和2.1倍，M-CSF分别为1.2倍和1.4倍。在这些不同条件下成熟的巨噬细胞表现出相似的CD14、CD64、CD71、HLA-DR和Max 1抗原表达，以及对U937细胞相似的体外抗肿瘤活性。在细胞因子存在的情况下进行培养可实现用于过继免疫治疗试验的活化巨噬细胞的大规模生产。
