Lambré C R, Greffard A, Gattegno L, Saffar L
INSERM, U 139, Hôpital H. Mondor, Créteil, France.
Immunol Lett. 1990 Jan;23(3):179-82. doi: 10.1016/0165-2478(90)90188-v.
Human mononuclear cells were isolated from peripheral blood by centrifugation over Ficoll Hypaque, followed by adherence to plastic dishes. Monocyte-derived macrophages were obtained after culture for 3 or 5 days of the adherent cells in RPMI medium containing 20% heat-inactivated foetal calf serum. The sialidase activities were assayed in the whole homogenate using sodium 4-methyl-umbelliferyl-alpha-D-neuraminate as substrate, at various pHs, ranging from 3.6 to 6. The in vitro differentiation of monocytes into macrophages from day 0 up to day 5 was accompanied by a significant (P less than or equal to 0.01) increase in the sialidase activity on both a per-cell (+360%) and a per-mg protein in the homogenate (+125%) basis.
通过在Ficoll Hypaque上离心从外周血中分离出人类单核细胞，随后使其贴附于塑料培养皿上。将贴壁细胞在含有20%热灭活胎牛血清的RPMI培养基中培养3天或5天后获得单核细胞衍生的巨噬细胞。使用4-甲基伞形酮基-α-D-神经氨酸钠作为底物，在3.6至6的不同pH值下，对全匀浆中的唾液酸酶活性进行测定。从第0天到第5天，单核细胞在体外分化为巨噬细胞的过程中，唾液酸酶活性在每细胞基础上（增加360%）和匀浆中每毫克蛋白质基础上（增加125%）均显著（P≤0.01）增加。
